ПОЛИПЕПТИД (ВАРИАНТЫ), ДЕЙСТВУЮЩИЙ В КОМПЛЕКСЕ С РЕЦЕПТОРОМ АНДРОГЕНА, НУКЛЕИНОВАЯ КИСЛОТА (ВАРИАНТЫ), ВЕКТОР (ВАРИАНТЫ) И КЛЕТКА-ХОЗЯИН (ВАРИАНТЫ)

Изобретение относится к биотехнологии и генной инженерии. Полипептид с последовательностью SEQ ID NO:3 связывается с рецептором андрогенов и увеличивает способность рецептора андрогенов трансактивировать андроген-чувствительный ген. Предложены также нуклеиновая кислота, кодирующая такой полипептид, и вектор, содержащий такую нуклеиновую кислоту. Группа изобретений может быть использована для создания трансгенных животных, экспрессирующих ген белка, действующего в комплексе с рецептором андрогена, служащих моделями для разработки лекарственных средств для лечения рака. 12 н. и 12 з.п. ф-лы.

АНТИСМЫСЛОВЫЕ ОЛИГОНУКЛЕОТИДЫ (ODN) К Smad7 И ИХ ПРИМЕНЕНИЕ В ОБЛАСТИ МЕДИЦИНЫ

Изобретение относится к области молекулярной биологии и может быть использовано в медицине. Получены химически модифицированные антисмысловые олигонуклеотиды (ODN), направленные на сайт 403 мРНК Smad7, которые отличаются тем, что in vivo демонстрируют существенно более высокие уровни биологической (ингибиторной) активности, чем в системе in vitro, и при этом не обнаруживают явных побочных эффектов. Предлагается использовать новые модифицированные формы ODN к Smad7 в качестве терапевтических агентов, в частности при лечении хронических воспалительных заболеваний пищеварительного тракта, таких как болезнь Крона и язвенные колиты. 3 н. и 3 з.п. ф-лы, 6 ил., 4 табл.

НОВЫЕ БИЦИКЛИЧЕСКИЕ НУКЛЕОЗИДЫ И ОЛИГОМЕРЫ, ПОЛУЧЕННЫЕ ИЗ НИХ

Изобретение относится к новым соединениям формулы (I) и олигонуклеотидам, полученным из них. В частности, настоящее изобретение относится к соединению формулы (I) где один из T1 и Т2 представляет собой OR1 или OR2; и другой из T1 и Т2 представляет собой OR1 или OR2; где R1 представляет собой Н или гидроксил-защитную группу, где указанная гидроксил-защитная группа независимо в каждом случае выбрана из ацетила, бензила, трет-бутилдиметилсилила, трет-бутилдифенилсилила, тритила, 4-монометокситритила, 4,4'-диметокситритила (DMTr), 4,4',4''-триметокситритила (TMTr), 9-фенилксантин-9-ила (пиксила) и 9-(п-метоксифенил)ксантин-9-ила (МОХ), и R2 представляет собой фосфорсодержащий фрагмент, где указанный фосфорсодержащий фрагмент представляет собой фосфорамидитный фрагмент, представленный формулой (X) где R5 представляет собой С1-С9 алкил, замещенный циано; R6 и R7 независимо представляют собой С1-С9 алкил; или совместно с атомом азота, к которому они присоединены, образуют гетероциклическое кольцо ...

ИММУНОСТИМУЛИРУЮЩИЕ ОЛИГОНУКЛЕОТИДЫ

Изобретение относится к области биотехнологии. Описана группа изобретений, включающая иммуностимулирующий олигонуклеотид, стимулирующий толл-подобный рецептор 21 (TLR21), вакцина для предупреждения или лечения болезни Ньюкасла, вектор экспрессии, иммуностимулирующая композиция для стимуляции толл-подобного рецептора 21 (TLR21), способ стимуляции толл-подобного рецептора 21 (TLR21) и способ вызывания иммунного ответа, опосредованный толл-подобным рецептором 21 (TLR21). В одном из вариантов реализации иммуностимулирующий олигонуклеотид содержит по меньшей мере один CpG мотив и обогащенную гуаниновыми нуклеотидами последовательность. Изобретение расширяет арсенал средств, стимулирующих TLR21. 6 н. и 1 з.п. ф-лы, 79 ил., 65 табл., 10 пр.

СПОСОБ ВЫПОЛНЕНИЯ МАНИПУЛЯЦИИ С ПОСЛЕДОВАТЕЛЬНОСТЬЮ НУКЛЕИНОВОЙ КИСЛОТЫ, УСТРОЙСТВО ДЛЯ ПРОВЕДЕНИЯ МАНИПУЛЯЦИЙ НА ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ, ПРОТОЧНЫЙ СОСУД, ТВЕРДЫЙ НОСИТЕЛЬ ДЛЯ ИММОБИЛИЗАЦИИ ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ, ТВЕРДЫЙ НОСИТЕЛЬ, СПОСОБ ПОЛУЧЕНИЯ НОСИТЕЛЯ ДЛЯ ИММОБИЛИЗАЦИИ ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ

Способ применим, в частности для анализа медицинских проб. Способ проведения манипуляции последовательностью нуклеиновой кислоты предусматривает обеспечение системы твердого носителя, с которым связан одноцепочечный олигонуклеотид, комплементарный специфической последовательности на нуклеиновой кислоте-мишени, более длинной, чем данный олигонуклеотид, добавление источника одноцепочечной нуклеиновой кислоты-мишени к системе твердого носителя, гибридизацию нуклеиновой кислоты-мишени с олигонуклеотидом и проведение манипуляции на гибридизованной нуклеиновой кислоте-мишени. Манипуляция может представлять собой, например, копирование или амплификацию последовательности нуклеиновой кислоты-мишени. В предпочтительном варианте обеспечен носитель в проточном сосуде, который облегчает промывание носителя для удаления примесей и оставления "чистой" пробы нуклеиновой кислоты-мишени, на которой может быть проведена манипуляция. Предпочтительно носитель имеет силоксановый матрикс, с которым связан олигонуклеотид ...

УЛУЧШЕННАЯ ОЛИГОНУКЛЕОТИДНАЯ КОНСТРУКЦИЯ ТИПА НАНОЧАСТИЦЫ, ОБЛАДАЮЩАЯ ВЫСОКОЙ ЭФФЕКТИВНОСТЬЮ, И СПОСОБ ЕЕ ПОЛУЧЕНИЯ

Изобретение относится к области биотехнологии, конкретно к самособирающейся в наночастицу олигонуклеотидной конструкции, и может быть использовано в медицине. Олигонуклеотидная конструкция согласно настоящему изобретению может быть полезной в качестве системы доставки на основе олигонуклеотида нового типа, а также инструмента для лечения злокачественных заболеваний, инфекционных заболеваний и т.п. Полимерное соединение, входящее в состав конструкции, связано с антисмысловым олигонуклеотидом посредством ковалентной связи для улучшения стабильности in vivo олигонуклеотида и эффективности его доставки в целевую клетку. В сравнении с известными аналогами заявленная олигонуклеотидная конструкция оптимизирована в отношении гомогенного материала, упрощена схема ее твердофазного синтеза, а размер двухцепочечной конструкции олиго-РНК может быть точно отрегулирован посредством контроля числа повторов блока и гидрофильного материала. 8 н. и 21 з.п. ф-лы, 18 ил., 4 табл., 6 пр.

АНТИСМЫСЛОВЫЕ НУКЛЕИНОВЫЕ КИСЛОТЫ

Изобретение относится к биохимии. Описан антисмысловой олигомер, который вызывает пропуск 50-го экзона в гене дистрофина человека, состоящий из нуклеотидной последовательности, комплементарной любой из нуклеотидных последовательностей, состоящих из 106-го - 126-го, 107-го - 127-го, 108-го - 127-го, 108-го - 128-го или 109-го - 129-го нуклеотидов, считая от 5'-конца 50-го экзона гена дистрофина человека. Также описана фармацевтическая композиция для лечения мышечной дистрофии, включающая в качестве действующего ингредиента антисмысловой олигомер или его фармацевтически приемлемую соль или гидрат. Изобретение расширяет арсенал средств для лечения мышечной дистрофии. 2 н. и 10 з.п. ф-лы, 31 ил., 15 табл., 54 пр.

СТАБИЛЬНЫЕ И РАСТВОРИМЫЕ АНТИТЕЛА, ИНГИБИРУЮЩИЕ VEGF

Изобретение относится к области биохимии, в частности к выделенной молекуле нуклеиновой кислоты, кодирующей рекомбинантное антитело или его фрагмент, где рекомбинантное антитело или его фрагмент нейтрализует VEGF человека, а также к экспрессирующему вектору и клетке-хозяину, ее содержащей. Изобретение позволяет эффективно экспрессировать антитело или его фрагмент, которое нейтрализует VEGF человека. 3 н. и 2 з.п. ф-лы, 15 ил., 12 табл., 7 пр.

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Изобретение относится к применимому в медицине соединению, содержащему модифицированный олигонуклеотид и конъюгирующую группу, при этом модифицированный олигонуклеотид состоит из 12-30 связанных нуклеозидов и содержит последовательность азотистых оснований, включающую часть из по меньшей мере 8 смежных азотистых оснований, комплементарную равной по длине части SEQ ID NO: 1, при этом последовательность азотистых оснований модифицированного олигонуклеотида по меньшей мере на 90% комплементарна SEQ ID NO: 1, где конъюгирующая группа содержитгде конъюгирующая группа связана с модифицированным олигонуклеотидом на 5'-конце или 3'-конце модифицированного олигонуклеотида. Предложено новое соединение и композиция на его основе, эффективные для производства лекарственного средства для лечения заболевания, опосредованного НВV. 4 н. и 36 з.п. ф-лы, 113 пр., 108 табл.

СОПРЯЖЕННЫЕ АНТИСМЫСЛОВЫЕ СОЕДИНЕНИЯ И ИХ ПРИМЕНЕНИЕ

Изобретение относится к области биотехнологии и медицине. Предложено соединение, сопряженное с тремя N-ацетилгалактозаминовыми лигандами. Соединение имеет структуру XXVI:где Тпредставляет собой группу, содержащую нуклеозид, нуклеотид, мономерную субъединицу, реакционноспособный сложный эфир, линкер, расщепляемый фрагмент или олигомерное соединение. Соединение обладает улучшенной активностью в клетках печени in vivo и пригодно для применения в способе лечения метаболического расстройства или в способе лечения сердечно-сосудистого расстройства. 15 з.п. ф-лы, 120 табл., 112 пр.

МОДУЛЯЦИЯ ЭКСПРЕССИИ ФАКТОРА 11

Изобретение относится к области биотехнологии. Описаны антисмысловые соединения, способы снижения уровня фактора 11 и способы лечения или предупреждения тромбоэмболических осложнений у индивидуума, нуждающегося в этом. Примерами патологических состояний, которые могут быть подвергнуты лечению путем введения антисмысловых соединений, нацеленных на фактор 11, являются тромбоз, эмболия и тромбоэмболия, такие как тромбоз глубокой вены, эмболия легких, инфаркт миокарда и инсульт. Антисмысловые соединения, нацеленные на фактор 11, могут быть также использованы в качестве профилактических средств для устранения риска развития у индивидуумов тромбоза и эмболии. 5 н. и 52 з.п. ф-лы, 169 табл., 47 пр.

НОВЫЕ МОНОМЕРЫ И ОЛИГОМЕРЫ ПЕПТИДНЫХ НУКЛЕИНОВЫХ КИСЛОТ

Изобретение относится к соединению общей формулы (I):в которой K представляет собой активную группу эфира карбоновой кислоты или -О-R; где Rобозначает атом Н, метальную, этильную, бензильную или трет-бутильную группу; Pr представляет собой атом Н или аминозащитную группу; # обозначает асимметричный атом С; Е представляет собой аденинильную, цитозинильную, псевдо-изоцитозинильную, гуанинильную, тиминильную, урацилильную или фенильную группу, при необходимости замещенную защитной группой для нуклеотидного основания; Rобозначает группу общей формулы (II):в которой Rобозначает группу эфира фосфоновой кислоты или группу фосфоновой кислоты; Rобозначает аминозащитную группу; m обозначает 1, 2, 3 или 4; и h обозначает 0, 1, 2 или 3; при условии что сумма m и h в общей формуле (II) находится в пределах: 2≤х≤5. Также предложены соединение формулы (VI), такое как указано в формуле изобретения, и фармацевтическая композиция. Предложенные соединения формулы (I) представляют новые мономеры для получения ...

СПОСОБ СИНТЕЗА НУКЛЕОЗИДНЫХ АНАЛОГОВ

Изобретение относится к способу получения 2',3'-О-алкилиден-β -нуклеозидного аналога, включающему взаимодействие 2', 3'-О-алкилиден β-фуранозилгалогенида формулы 1, где R3 и R4 представляют собой водород или (CH2)nQ, где n является целым числом от 1 до 6; Q представляют собой водород или -OR7; R5 и R6 представляют собой (С1-С6) алкил; R7 представляет собой водород или защитную группу; Y представляет хлор или бром, с гетероциклом формулы 2, где каждый из R8 представляет собой водород, фенил, фенил, замещенный галогеном, OR7 или NR7R7; R7 представляет собой водород, фенил, фенил, замещенный галогеном или защитную группу. Реакцию проводят в диметилсульфоксиде или в смеси полярных растворителей, в которой объем диметилсульфоксида составляет, по крайней мере, примерно 15%, в присутствии сильного основания, представляющего собой т-бутоксид натрия или т-бутоксид калия. Молярное отношение 2',3'-O-алкилиден-β-фуранозилгалогенида к гетероциклу составляет около (1:1) - (3:1). Молярное отношение сильного ...

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ АПОЛИПОПРОТЕИНА C-III

Изобретение относится к пригодному для применения в медицине соединению, содержащему модифицированный олигонуклеотид и конъюгирующую группупри этом модифицированный олигонуклеотид состоит из 20 связанных азотистых оснований, комплементарных равной по длине части азотистых оснований 3533-3552 в SEQ ID NO: 3, при этом модифицированный олигонуклеотид содержит сегмент гэп, состоящий из связанных дезоксинуклеозидов; сегмент крыла 5', состоящий из связанных нуклеозидов; сегмент крыла 3', состоящий из связанных нуклеозидов; при этом сегмент гэп расположен между сегментом крыла 5' и сегментом крыла 3', и при этом каждый нуклеозид каждого сегмента крыла содержит модифицированный сахар, и где по меньшей мере один нуклеозид содержит модифицированное азотистое основание; и модифицированный олигонуклеотид содержит по меньшей мере одну модифицированную межнуклеозидную связь. Предложены новые конъюгаты модифицированных олигонуклеотидов и композиции на их основе для лечения, предупреждения или замедления ...

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ HBV И TTR

Группа изобретений относится к области фармакологии и медицины и направлена на лечение транстиретинового амилоидоза. Раскрывается олигомерное соединение, при этом анионная форма олигомерного соединения представлена следующей химической структурой (SEQ ID NO: 12). Кроме того, раскрыта форма свободной кислоты, натриевая соль вышеуказанного олигомерного соединения и олигомерное соединение, содержащее модифицированный олигонуклеотид и конъюгирующую группу, характеризующееся следующей формулой (SEQ ID NO: 12). Также раскрыта фармацевтическая композиция для лечения транстиретинового амилоидоза на основе вышеуказанных соединений, применение представленных соединений при производстве лекарственного средства для лечения транстиретинового амилоидоза, а кроме того, способ лечения транстиретинового амилоидоза у субъекта, включающий введение субъекту терапевтически эффективного количества олигомерного соединения или фармацевтической композиции, содержащей указанное олигомерное соединение. Группа изобретений ...

Алкинсодержащий амидофосфит для функционализации синтетических олигонуклеотидов и способ его получения

Группа изобретений относится к области органической химии и химии нуклеиновых кислот и предназначена для функционализации синтетических олигонуклеотидов методом азид-алкинового циклоприсоединения («клик-химии»). Раскрывается соединение, имеющее структурную формулу, где R1 представляет собой защитную группу, стабильную в условиях синтеза олигонуклеотидов, выбранную из 4,4'-диметокситритильной (DMTr), 4-монометокситритильной (MMTr), тритильной (Tr), триметокситритильной (TMTr), 9-фенилксантен-9-ильной (Px), карбонатной или силильной групп, R2 представляет собой водород, метил или этил, X представляет собой -О- или -NH-, и L представляет собой спейсерную группу -(CH2)-n, где n = 1, 2 или 3; или -(CH2CH2O)nCH2-, где n = 1 или 2. Также раскрыт способ получения вышеуказанных амидофосфитных реагентов, включающий функционализацию карбоксильной группы линкерами, содержащими алкиновую группу, с последующей селективной защитой одной гидроксильной группы и фосфитилированием второй гидроксильной группы ...

КОМПОЗИЦИИ И СПОСОБЫ КОНТРОЛЯ ВРЕДИТЕЛЕЙ РАСТЕНИЙ

Изобретение относится к области биохимии, в частности к химерной генетической конструкции для экспрессии белка, токсичного для совки-ипсилон (Agrotis ipsilon), а также к содержащему ее вектору, клетке, растению, семени растения. Также раскрыта синтетическая молекула нуклеиновой кислоты для обеспечения экспрессии белка, токсичного для совки-ипсилон (Agrotis ipsilon), а также способ получения устойчивого к совке-ипсилон трансгенного растения. Изобретение также относится к белку, токсичному для совки-ипсилон (Agrotis ipsilon), к способу его получения, а также к содержащей его композиции и к способу борьбы с совкой-ипсилон (Agrotis ipsilon) с его использованием. 11 н. и 16 з.п. ф-лы, 3 табл., 11 пр.

КОМПОЗИЦИЯ ДЛЯ ЛЕЧЕНИЯ СОСТОЯНИЙ, ЧУВСТВИТЕЛЬНЫХ ИЛИ ВОСПРИИМЧИВЫХ К ВОЗДЕЙСТВИЮ ЛОЖНОСПАРЕННОЙ DS РНК

Изобретение относится к медицине. Цель - повышение стабильности. Композиция содержит носитель, ложноспаренную dsРНК, поверхностно-активное вещество в определенных количествах. Ложноспаренная dsРНК содержит участки разрыва связи и имеет формулу rIn•r (C11-14• V)n, где n - целое число от 4 до 29. 2 табл.

ОЛИГОМЕРНЫЕ СОЕДИНЕНИЯ ДЛЯ МОДУЛЯЦИИ BCL-2

... 1. Олигомерное соединение, имеющее длину 10-30 нуклеиновых оснований и содержащее мишень-связывающий домен, который специфически гибридизуется с областью от положения основания 1459 (5') до 1476 (3') мРНК человеческого Bcl-2 (HUMBcl2A (номер доступа М13994) в базе данных GenBank), где указанный связывающийся с мишенью домен имеет формулу 5'-[(ДНК/РНК)0-1-(LNA/LNA*)2-7-(ДНК/РНК/LNA*)4-14-(LNA/LNA*)2-7-(ДНК/РНК)0-1]-3', где LNA означает нуклеотид LNA, LNA* означает нуклеотидный аналог LNA; и указанный мишень-связывающий домен содержит, по меньшей мере, два нуклеотида LNA или нуклеотидных аналога LNA, связанных фосфортиоатной группой (-О-Р(О,S)-О-). 2. Соединение по п.1, где, по меньшей мере, 70% нуклеотидных связей в указанном мишень-связывающем домене представляют собой фосфортиоатные группы (-О-Р(О,S)-О-). 3. Соединение по п.2, где все нуклеотидные связи в олигомерном соединении представляют собой фосфортиоатные группы. 4. Соединение по п.1, где 10-50% нуклеиновых оснований в мишень-связывающем ...

СПОСОБ ПОЛУЧЕНИЯ АЛКИЛЗАМЕЩЕННЫХ 2-ДЕКОКСИ-2-ФТОР-D-РИБОФУРАНОЗИЛ-ПИРИМИДИНОВ И ПУРИНОВ И ИХ ПРОИЗВОДНЫХ

... 1. 3,5-ди-О-защищенный 2-дезокси-2-фтор-2-С-метил-D-рибоно-γ-лактон общей формулы: где R3 и R5 могут независимо обозначать Н, СН3 или ацил; или R3 и R5 соединены через -SiR2-O-SiR2- или -SiR2 -, где R обозначает низший алкил, такой как СН3, этил и н-Pr или изо-Pr. 2. Соединение по п.1, отличающееся тем, что R3 и R5 каждый независимо обозначает Н, СН3, ацетил, бензоил, пивалоил, 4-нитробензоил, 3-нитро-бензоил, 2-нитробензоил, 4-хлорбензоил, 3-хлорбензоил, 2-хлорбензоил, 4-метилбензоил, 3-метилбензоил, 2-метилбензоил, 4-фенилбензоил, бензид, 4-метоксибензил, тритил, триалкилсилил, трет.бутилдиалкилсилил, трет.бутилдифенилсилил, TIPDS, ТНР, MOM или MEM. 3. Способ получения лактона по п.1, включающий стадии: (а) реакции соединения формулы 39 с алкил-2-бромпропионатом в присутствии активированного цинка в подходящем растворителе; (b) добавления окисляющего агента с получением кетона; (c) фторирования продукта со стадии (b) с получением фторированного кетона; (d) восстановления фторированного ...

БИОЛОГИЧЕСКИ АКТИВНОЕ СОЕДИНЕНИЕ, СОДЕРЖАЩЕЕ КОДИРУЮЩИЙ ОЛИГОНУКЛЕОТИД (ВАРИАНТЫ), СПОСОБ ЕГО СИНТЕЗА, БИБЛИОТЕКА СОЕДИНЕНИЙ (ВАРИАНТЫ), СПОСОБ ЕЕ СИНТЕЗА, И СПОСОБ ПОИСКА СОЕДИНЕНИЯ, СВЯЗЫВАЮЩЕГОСЯ С БИОЛОГИЧЕСКОЙ МИШЕНЬЮ (ВАРИАНТЫ)

... 1. Способ синтеза соединения, содержащего функциональный компонент, который эффективно связан с кодирующим олигонуклеотидом, отличающийся тем, что в качестве исходного соединения используют соединение-инициатор, состоящее из исходного функционального компонента, содержащего n структурных фрагментов, где n равно целому числу 1 или более, по крайней мере одну реакционноспособную группу, и эффективно связанного с исходным олигонуклеотидом; осуществляют взаимодействие указанного соединения-инициатора со структурным фрагментом, содержащим по крайней мере одну реакционноспособную группу, комплементарную реакционноспособной группе соединения-инициатора, в условиях, пригодных для образования ковалентной связи между реакционноспособными группами; осуществляют взаимодействие исходного олигонуклеотида с новым вводимым олигонуклеотидом, который определяет структурный фрагмент, ковалентно связанный с соединением инициатором, причем взаимодействие исходного олигонуклеотида с новым олигонуклеотидом осуществляют ...

АНТИТЕЛА ПРОТИВ РЕЦЕПТОРА ФОЛИЕВОЙ КИСЛОТЫ 1, ИХ ИММУНОКОНЪЮГАТЫ И ИСПОЛЬЗОВАНИЕ

... 1. Гуманизированное антитело или его антиген-связывающий фрагмент, специфически связывающиеся с человеческим рецептором фолиевой кислоты 1, содержащее:(а) CDR1 тяжелой цепи, содержащую GYFMN (SEQ ID NO:1); CDR2 тяжелой цепи, содержащую RIHPYDGDTFYNQXaaFXaaXaa(SEQ ID NO:56); и CDR3 тяжелой цепи, содержащую YDGSRAMDY (SEQ ID NO:3); и(б) CDR1 легкой цепи, содержащую KASQSVSFAGTSLMH (SEQ ID NO:7); CDR2 легкой цепи, содержащую RASNLEA (SEQ ID NO:8); и CDR3 легкой цепи, содержащую QQSREYPYT (SEQ ID NO:9);где Xaaвыбран из K, Q, Н и R; Хаавыбран из Q, И, N и R; и Хаавыбран из G, Е, T, S, A и V.2. Гуманизированное антитело или его антиген-связывающий фрагмент по п.1, отличающиеся тем, что последовательность CDR2 тяжелой цепи содержит RIHPYDGDTFYNQKFQG (SEQ ID NO:2).3. Гуманизированное антитело или его антиген-связывающий фрагмент по п.1 содержащие вариабельный домен тяжелой цепи, по меньшей мере приблизительно на 90%, приблизительно на 95%, приблизительно на 99%, приблизительно на 100% идентичный ...

Носитель для твердофазного синтеза, способ его получения и его применение

Изобретение относится к области получения функциональных полимерных материалов, а именно, представляет собой носитель для использования в твердофазном синтезе олигонуклеотидов. Носитель для твердофазного синтеза описывается формулой: где R1= -(CH2)n-, n – целое число о 0 до 3, или R1= -O-(CH2)m-O-, m – целое число от 1 до 4; и R2= -OH или -NH2; при этом получение носителя для твердофазного синтеза включает следующие этапы: вначале получают водную и масляную фазы; водная фаза содержит воду, диспергирующее средство и неорганическую соль; масляная фаза содержит: поперечно-сшивающий мономер, моновинильное соединение, функциональный мономер, модифицированный мономер; порообразующий агент и инициатор. Затем масляную фазу добавляют к водной при перемешивании и нагревании для проведения реакции полимеризации, после чего, удаляют порообразующий агент. В результате получают пористую полимерную смолу, которая способна подвергаться дальнейшей реакции для получения носителя твердофазного синтеза, содержащего ...

СПОСОБ СИНТЕЗА В РАСТВОРЕ МАКРОМОЛЕКУЛ ИЗ ЗВЕНЬЕВ ПРОИЗВОДНЫХ УГЛЕВОДОВ

Изобретение относится к синтезу макромолекул, состоящих из моносахаридных или олигосахаридных звеньев. Предложен способ синтеза макромолекул, состоящих из звеньев, представляющих собой моносахариды или производные моносахаридов, путём последовательного удлинения цепи на мономер или олигомер, обладающий по меньшей мере двумя функциональными группами, где на первой стадии происходит заякоривание мономера или концевого звена олигомера якорной молекулой с последующим удалением защитной группы, оставляя свободную функциональную группу, к которой затем присоединяется второй мономер или олигомер, который может содержать свободную функциональную группу, защищённую защитной группой, при этом указанная якорная молекула содержит полиолефиновую цепь, или полиолефиновый олигомер, или полиалкен с по меньшей мере от 10 до 50 мономерными звеньями, указанная полиолефиновая цепь представляет собой разветвленную цепь и предпочтительно полиизобутеновую цепь; а также применение указанного способа для синтеза ...

СТАБИЛЬНЫЕ И РАСТВОРИМЫЕ АНТИТЕЛА, ИНГИБИРУЮЩИЕ VEGF

Изобретение относится к области иммунологии и биотехнологии. Представлены вариантные рекомбинантные антитела против VEGF человека, имеющие вариабельные области тяжелой и легкой цепей, содержащие гипервариабельне участки (CDR) антител кролика. Также представлены: выделенные молекулы нуклеиновых кислот, кодирующие указанные антитела; вектор экспрессии, содержащий указанную молекулу нуклеиновой кислоты; и клетка-хозяин для экспрессии антитела по изобретению, содержащая указанный вектор экспрессии. Описана фармацевтическая композиция, содержащая терапевтически эффективное количество указанного антитела и фармацевтически приемлемый носитель. Изобретение позволяет расширить арсенал антител против VEGF человека, полученных от кролика. 7 н. и 17 з.п. ф-лы, 15 ил., 12 табл., 7 пр.

УЛУЧШЕННАЯ ОЛИГОНУКЛЕОТИДНАЯ КОНСТРУКЦИЯ ТИПА НАНОЧАСТИЦЫ, ОБЛАДАЮЩАЯ ВЫСОКОЙ ЭФФЕКТИВНОСТЬЮ, И СПОСОБ ЕЕ ПОЛУЧЕНИЯ

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ ПКП

СПОСОБ ПОЛУЧЕНИЯ СОЕДИНЕНИЯ, СОДЕРЖАЩЕГО ФУНКЦИОНАЛЬНЫЙ КОМПОНЕНТ И ОЛИГОНУКЛЕОТИД (ВАРИАНТЫ), ВЫШЕУКАЗАННОЕ СОЕДИНЕНИЕ (ВАРИАНТЫ), СПОСОБ СИНТЕЗА БИБЛИОТЕКИ СОЕДИНЕНИЙ, БИБЛИОТЕКА СОЕДИНЕНИЙ (ВАРИАНТЫ), СПОСОБ ИДЕНТИФИКАЦИИ СОЕДИНЕНИЯ, СВЯЗЫВАЮЩЕГОСЯ С БИОЛОГИЧЕСКОЙ МИШЕНЬЮ (ВАРИАНТЫ)

... 1. Способ получения соединения, содержащего функциональный компонент, который эффективно связан с кодирующим олигонуклеотидом, в котором в качестве исходного соединения используют соединение-инициатор, которое состоит из исходного функционального компонента, эффективно связанного с исходным олигонуклеотидом, при этом исходный функциональный компонент содержит n структурных фрагментов, где n равно целому числу 1 или более, и по крайней мере одну реакционноспособную группу, осуществляют взаимодействие указанного соединения-инициатора со структурным фрагментом, содержащим по крайней мере одну реакционноспособную группу, которая комплементарна по меньшей мере одной реакционноспособной группе исходного функционального компонента, в условиях, пригодных для образования ковалентной связи между комплементарными реакционноспособными группами, вводят новый олигонуклеотид, определяющий структурный фрагмент, и осуществляют взаимодействие исходного олигонуклеотида с вводимым олигонуклеотидом в присутствии ...

АНАЛОГИ ОЛИГОНУКЛЕОТИДОВ КАК СРЕДСТВА КОРРЕКЦИИ СПЛАЙСИНГА ДЛЯ ЛЕЧЕНИЯ МЫШЕЧНОЙ ДИСТРОФИИ ДЮШЕНА

ХИМЕРНЫЕ РЕЦЕПТОРЫ АНТИГЕНА, НАЦЕЛЕННЫЕ НА PSCA

СПОСОБ СПЕЦИФИЧЕСКОГО ВЫДЕЛЕНИЯ ПОЛНОГО ДНК-СОДЕРЖИМОГО БАКТЕРИАЛЬНЫХ ВОБУДИТЕЛЕЙ ИНФЕКЦИИ И НАБОР, ИСПОЛЬЗУЮЩИЙСЯ ДЛЯ ЭТИХ ЦЕЛЕЙ

... 1. Способ специфического выделения полного ДНК-содержимого бактериальных возбудителей инфекции из образцов, в ходе которого клетки лизируют, ДНК-содержимое лизата выборочно связывают, промывают и затем обессоленную линейную полимерную нуклеиновую кислоту элюируют со связывающей поверхности водным раствором, отличающийся тем, что на предварительном этапе нежизнеспособные клетки отделяют от жизнеспособных на основе различных физико-химических свойств их клеточной поверхности, различной проницаемости поверхности этих клеток и аффинитета ионов, используя промывочный раствор с различной проникающей способностью, содержащий детергент и электролит, предпочтительно буферную смесь из Тритона Х-100, этилендиаминтетрауксусной кислоты, KCl или NaCl, моногидрата лимонной кислоты; при этом изменяют центробежную силу, влияющую на осаждение клеток, в результате чего только двухцепочечные ДНК, полученные при лизисе жизнеспособных клеток, связывают с матрицей -SiO-TiO-, содержащей химически активированные ...

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ HBV И TTR

ВАРИАНТЫ ОСУЩЕСТВЛЕНИЯ ЗОНДА И СПОСОБЫ НАПРАВЛЕННОГО ДЕЙСТВИЯ НА НУКЛЕИНОВЫЕ КИСЛОТЫ

... 1. Двухцепочечный зонд, включающийпару мономеров, включающую первый мономер, имеющий формулугде каждый Y независимо выбран из углерода, кислорода, серы, триазола, оксазола, тетразола, изоксазола и NR, где Rвыбран из водорода, алифатического соединения, арила, гетероалифатического соединения и гетероарила; Rи Rвыбраны из водорода, алифатического соединения, арила, алифатического арила и части, содержащей гетероатом; или Rвыбран из функциональной группы, содержащей гетероатом; Rпредставляет собой функциональную группу, содержащую гетероатом; Rвыбран из любого природного или неприродного нуклеинового основания; Rвыбран из интеркалятора, подходящего для интеркаляции в нуклеиновую кислоту; "необязательный линкер" выбран из линкеров, включающих алкильные линкеры, амидные линкеры, карбаматные линкеры, карбонатные линкеры, мочевинные линкеры и их сочетания;второй мономер, имеющий формулугде Y, R, R, R, R, Rи "необязательный линкер" представляют собой указанное для первого мономера; V выбран из ...

Модифицированный биосенсор для детекции внутриклеточной РН

... 1. Нуклеиновая кислота, кодирующая флуоресцентный биосенсор для регистрации изменения рН, аминокислотная последовательность которого показана в SEQ ID No: 4.2. Кассета экспрессии, которая будучи интегрированной в геном клетки или при введении в клетку в виде внехромосомного элемента, способна обеспечить экспрессию флуоресцентного биосенсора по п. 1 под контролем регуляторных элементов, необходимых для экспрессии нуклеиновой кислоты в клетке-хозяине.3. Эукариотическая клетка, продуцирующая биосенсор, кодируемый нуклеиновой кислотой по п. 1, содержащая кассету экспрессии по п. 2 в виде внехромосомного элемента или элемента, интегрированного в геном этой клетки.4. Нуклеиновая кислота, кодирующая химерный белок, аминокислотная последовательность которого выбрана из SEQ ID NOs:6 и 8, где химерный белок включает флуоресцентный биосенсор по п. 1.5. Кассета экспрессии, которая будучи интегрированной в геном клетки или при введении в клетку в виде внехромосомного элемента, способна обеспечить экспрессию ...

МОДУЛЯЦИЯ ЭКСПРЕССИИ ФАКТОРА 11

... 1. Соединение, включающее модифицированный олигонуклеотид, состоящий из 12-30 связанных нуклеозидов, нацеленное на нуклеиновую кислоту фактора 11, в котором нуклеотидная последовательность модифицированного олигонуклеотида по меньшей мере на 90% комплементарна части последовательности нуклеиновой кислоты фактора 11, имеющей такую же длину, где необязательно указанная нуклеиновая кислота фактора 11 представляет собой нуклеиновую кислоту фактора 11, представленную в SEQ ID NO: 1 или SEQ ID NO: 2.2. Соединение по п. 1, в котором модифицированный олигонуклеотид включает нуклеотдную последовательность, включающую часть по меньшей мере из 8 смежных нуклеотидных оснований, которая комплементарна имеющей такую же длину части:(i) нуклеотидных оснований 675-704 в SEQ ID NO: 1,(ii) нуклеотидных оснований 1062-1090 в SEQ ID NO: 1,(iii) нуклеотидных оснований 656-676 в SEQ ID NO: 1,(iv) нуклеотидных оснований 665-687 в SEQ ID NO: 1,(v) нуклеотидных оснований 738-762 в SEQ ID NO: 1,(vi) нуклеотидных ...

СОЕДИНЕНИЕ, СПОСОБ И ФАРМАЦЕВТИЧЕСКАЯ КОМПОЗИЦИЯ ДЛЯ МОДУЛИРОВАНИЯ ЭКСПРЕСИИ DUX4

Предложенная группа изобретений относится к области биотехнологии и может быть использована в медицине. Раскрываются модифицированные олигонуклеотиды, способные к ингибированию экспрессии гена двойного гомеобокса 4 (DUX4). Изобретение может быть применимо при лечении заболеваний, связанных аномальной экспрессий указанного гена у индивидуума, в частности при антисмысловой генной терапии фациоскапуло-плечевой мышечной дистрофии (FSHD), В-клеточного острого лимфоцитарного лейкоза, дифференцированной круглоклеточной саркомы и рабдомиосаркомы плода.10 н. и 19 з.п. ф-лы, 6 ил., 5 табл., 12 пр.

Способ получения синтетической двунитчатой РНК, обладающей интерферониндуцирующей активностью

VERFAHREN ZUR KONTROLLIERTEN SYNTHESE VON BELIEBIGEN PEPTIDMISCHUNGEN KODIERENDE POLYNUKLEOTIDMISCHUNGEN

ETIKETTIERREAGENZ UND NACHWEISMETHODE

7-DEAZAPURIN MODIFIZIERTE OLIGONUKLEOTIDE

VERFAHREN ZUR SCHNELLEN SYNTHESE VON GROSSEN ZAHLEN TRAEGERGEBUNDENER ODER FREIER PEPTIDE

The invention concerns a method for the rapid synthesis of substrate-bound or free peptides, apparatus for carrying out the method, a flat material with peptides prepared by the method bound to it, and a use of this material.

DNS-SEQUENZBESTIMMUNG DURCH MASSENSPEKTROMETRIE AUF DEM WEG DES ABBAUS MIT EXONUKLEASE

POLYNUKLEOTID REAGENZIEN MIT NICHTNUKLEOTIDISCHEN GRUPPIERUNGEN, DEREN HERSTELLUNG UND VERWENDUNG

OLIGO-2'-DESOXYNUKLEOTIDE UND IHRE VERWENDUNG ALS ARZNEIMITTEL MIT ANTIVIRALER WIRKSAMKEIT

The invention concerns oligodeoxyribonucleotides, in which, at both the 5' and the 3' terminal positions, at least two 2'-deoxy- beta -D-erythro-pentofuranosyl groups have been replaced by 2'-deoxy- beta -D-threo-pentofuranosyl groups, and oligodeoxyribonucleotides in which at least 20 % of the 2'-deoxy- beta -D-erythro-pentofuranosyl groups in successive nucleotide units have been replaced by 2'-deoxy- beta -D-threo-pentofuranosyl groups, and which consist of 6 to 100 nucleotide units. Such oligodeoxyribonucleotides are suitable for use as anti-sense inhibitors of the expression of viral genes and oncogenes and can be used in the preparation of drugs with an anti-viral action.

Markierte Oligonukleotide

VERFAHREN, TESTSÄTZE UND REAKTIVE TRÄGER ZUR 3'-MARKIERUNG VON OLIGONUKLEOTIDEN

2'-5'-Phosphorothioat-Oligoadenylate und deren antivirale Verwendungen

OLIGONUCLEOTID-POLYAMID KONJUGATE

DEM MENSCHLICHEN GEWEBEFAKTOR ÄHNLICHE DNS-SEGMENTE, POLYPEPTIDE UND ANTIKÖRPER

IN DER HAUPTKETTE MODIFIZIERTE OLIGONUKLEOTID-ANALOGE

ANTISENSE OLIGONUKLEOTIDE

2-AZAPURINE VERBINDUNGEN UND IHRE VERWENDUNG

Auf einer Oberfläche befestigte multifunktionelle Polymere-Netzwerke für Sensorchips

Hydrophobe Nukleinsäure-Sonde

NUKLEOSID-DERIVATE UND IHRE VERWENDUNG IN DER OLIGONUKLEOSID-SYNTHESE.

OLIGONUKLEOTIDE SYNTHESE

Verfahren zur Herstellung von N-Glykosiden

Effektive Gensonden-Konjugationen mittels eines aussergewöhnlichen Mischanhydridverfahrens

0,0'-Dicyanoethyl-phosphoramidite, Verfahren zu deren Herstellung und ihre Verwendung in Phosphorylierungsreaktionen.

Einschrittverfahren und Polynukleotidverbindungen zur Hybridisierung mit Ziel-Polynukleotiden

Markierte Nukleinsäuren.

2'-MODIFIZIERTE NUKLEOTIDE

Sonden

DEVICE FOR SIMULTANEOUSLY CARRYING OUT A PLURALITY OF CHEMICAL REACTION STEPS

Vorrichtung zum Nachweis von fluoreszenten, der Bindung eines Liganden auf einem Substrat entsprechenden Merkmalen

Therapeutische Anwendungen von 2',5'-Oligoadenylat-Derivaten

UNGELADENE, AUF MORPHOLIN BASIERENDE POLYMERE MIT CHIRALEN, PHOSPHOR ENTHALTENDEN BRÜCKEN ZWISCHEN DEN UNTEREINHEITEN

OLIGONUKLEOTID-ANALOGE.

ANTISENSE-MODULATION DER SURVIVIN-EXPRESSION

New 2-(1-((hetero)aryl(oxy))-ethyl)-nitrobenzene derivatives, useful for introducing intramolecular triplet sensitized photolabile protecting groups into synthons for light-controlled biopolymer production

... 2-(1-((Hetero)aryl or (hetero)aryloxy)-ethyl)-nitrobenzene derivatives (I) are new. Also new are nucleotide synthon compounds (II) containing a photolabile protecting group derived from (I). Nitrobenzene derivatives of formula (I) are new. R1 = halo, CN or NO2; or alkyl, alkenyl, alkynyl, alkoxy, alkoxycarbonyl, aryl, aryloxy, heteroaryl or heteroaryloxy (all optionally substituted, aryl or heteroaryl moieties being mono- or polycyclic); or adjacent groups R1 may together form a 5- or 6-membered ring (optionally containing heteroatoms); R2 = mono- or polycyclic aryloxy or heteroaryloxy; or polycyclic aryl or heteroaryl containing at least 3 fused rings; R3 = trialkylsilyl; or alkyl, alkenyl, alkynyl, alkoxy, alkoxycarbonyl, aryl, aryloxy, heteroaryl or heteroaryloxy (all optionally substituted, aryl or heteroaryl moieties being mono- or polycyclic); m = 0-4; p = 0 or 1; X = -C(O)-, -O-C(O)-, -O-C(S)- or -S(O)2-; and Y = H, leaving group or substrate. An Independent claim is included for ...

New 3'-derivatised oligo:nucleotide analogues

... 3'-Modified oligonucleotides (I) and (II) and their salts are new. In the formula, Q = gp. (a), i.e. cpds. (I), or gp. (b), i.e. cpds. (II); a,b = 0-20; R<1> = H; alkyl, alkenyl or alkynyl with up to 18C; 1-18C alkylcarbonyl; 2-19C alkenylcarbonyl; 3-19C alkynylcarbonyl; 6-20C aryl; 6-14C aryl-1-8C alkyl; or a gp. -P(W)(Z)(Z') (III); R<2> = H; OH; 1-18C alkoxy; halo; N-3; or NH2; D = OH or OPO3<2->; B = conventional nucleotide base linked in the alpha- or beta-configuration; n = 1-100; n' = 0-50; m = 0-5; m' = 0-5 in cpds. (I) and 1-5 in cpds. (II); A = O; S or CH2; W = O, S or Se; V = O or sulphanediyl; T = O; sulphanediyl; NH; or CH2; X = OH or SH; U = OH; SH; BH3; SeH; alkyl or alkoxy with up to 18C; 6-20C aryl; 6-14C aryl-1-8C alkyl; NHR<3>; NR<3>R<4>; or a gp. (OCH2CH2)pO(CH2)qCH2R<5> (IV); R3 = 1-18C alkyl; 6-20C aryl; 6-14C aryl-1-8C alkyl; or (CH2)c-NH(CH2)cd-NR<6>R<6>; and R<4> = 1-18C alkyl; 6-20C aryl; or 6-10C aryl-1-8C alkyl; or NR<3>R<4> forms a 5- or 6-membered heterocycle ...

SULFONATFREIE CYANINFARBSTOFFE ZUR MARKIERUNG VON NUKLEOSIDEN UND NUKLEOTIDEN

NEUE PSORALENDERIVATE

Massives Parallelverfahren zur Dekodierung von DNA und RNA

Ein Nukleotidanalogon, welches umfasst: a) eine Base ausgewählt aus der Gruppe bestehend aus Adenin oder einem Analogon von Adenin, Cytosin oder einem Analogon von Cytosin, Guanin oder einem Analogon von Guanin, Thymin oder einem Analogon von Thymin sowie Uracil oder einem Analogon von Uracil; b) eine eindeutige Markierung, welche mittels eines abspaltbaren Linkers an der Base oder an einem Analogon der Base befestigt ist; c) eine Desoxyribose; und d) eine abspaltbare chemische Gruppe, um eine -OH-Gruppe an einer 3'-Position der Desoxyribose zu schützen, wobei die abspaltbare chemische Gruppe durch chemische Mittel abgespalten werden kann.

DNA Chip, PNA Chip, sowie Herstellungsverfahren

FLT4 (VEGFR-3) ALS EIN ZIEL FÜR KREBSDARSTELLUNG UND ANTI-KREBS-BEHANDLUNG

ISOLIERUNG VON EINZELSTRÄNGIGEN NUKLEINSÄUREN

NANOPARTIKEL MIT DARAN ANGEHEFTETEN OLIGONUKLEOTIDEN SOWIE DEREN VERWENDUNGEN

FESTPHASEN-SYNTHESE

Energieübertragungsfarbstoffe mit verbesserter Fluoreszenz

Methoden zur Analyse von Nukleinsäure-Molekülen mittels Techniken zur Grössen-bestimmung

FORMULIERUNGEN UND VERFAHREN ZUM DENATURIEREN VON PROTEINEN

Zytostatisch wirksame Mittel

Neue chemische Verbindungen und ihre Darstellungen und Behandlungen

Programming nucleic and amino acids for computer calculation

Preparation method for the programming of substances (amino and nucleic acids) that act together in the genetic code, comprises assigning binary numbers to atomic weights and submolecular structures.

Filtration process

Signalling means employing lanthanide ions

Homogeneous assays for determining polynucleotide analytes are disclosed. The analyte is detected by a process wherein the binding of a complementary probe to the analyte allows the binding of an intercalating agent or a groove binder. The complementary probe and duplex binder carry different partners of a ligand-to-metal energy transfer system which comprises an energy donor and an energy acceptor. Figure 1 illustrates an embodiment wherein probe DNA is linked to a chelator e.g. EDTA, DTPA, DCTA and the intercalator is linked to a sensitiser. The chelator and sensitiser form a complex around a lanthanide ion e.g. europium, terbium, samarium, dysposium. The system is irradiated with light which can be selectively absorbed by the sensitiser and which can efficiently donate its energy to the lanthanide ion because the metal ion is chelated to the sensitiser. The delayed luminescence of the lanthanide ion marks the binding of the probe to the analyte. Also disclosed are duplex binders linked ...

Derivatives of 7-deaza-2'-deoxy-guanosine-5'-triphosphate, preparation and use thereof

Molecule comprising the following moiety: wherein R 1 is C 1-10 alkyl group optionally substituted by hydroxyl, amino, C 1-4 alkoxy or halo; and R 2 is hydrogen or hydroxyl and R 7 is H or a mono-, di- or tri-phosphate or thiophosphate thereof. These compounds are used to prepare oligonucleotide sequences containing the 7-deaza species and are useful in biological processes such as resolution of compression artifacts in DNA sequencing.

Compounds for mass spectrometry

Method for inhibiting viral propagation and anti-viral agent

Described is a method for inhibiting propagation of virus which comprises delivering one or more oligodeoxynucleotides or polydeoxynucleotides to a place where a messenger RNA (mRNA) generated on propagation of the virus exists, characterized in that the oligodeoxynucleotides or polydeoxynucleotides are identical in DNA sequence with a portion of the structure of DNA hybridizable to the mRNA, the structure being capable of hybridizing to the mRNA. Also described is an anti-viral agent which comprises an effective amount of one or more oligodeoxynucleotides or polydeoxynucleotides with a conventional liquid vehicle or excipient, characterized in that the oligodeoxynucleotides or polydeoxynucleotides are identical in DNA sequence with a portion of the structure of DNA hybridizable to an mRNA generated on propagation of the virus, the structure being capable of hybridizing to the mRNA. The method and the agent can widely be applied to the treatment of viral diseases.

Method for the preparation of a probe for nucleic acid hybridization

POLYNUCLEOTIDES AND THEIR PRODUCTION

... 1355516 Interferon-inducing medicines MERCK & CO Inc 10 May 1971 [15 May 1970] 14050/71 Heading A5B [Also in Division C3] In a polyriboinosinic-acid-polyribocytidylicacid complex having interferon-inducing activity, the polyribocytidylic acid component contains fewer than 100 nucleotide units. The polyriboinsinic acid component may have more than 500 nucleotide units. A complex rI n :rC n may be made according to Specification 1187830 and treated with sonic radiation to reduce the number of cytidylic nucleotide units. Alternatively, rC n homopolymers may be treated with sonic radiation or with ribonuclease to reduce the polymer to less than 100 units, and then complexed according to Specification 1,187,830 with rI n having more than 500 units. They may be administered intra-nasally or by injection.

PHARMACEUTICAL PREPARATION

5'-modified bicyclic nucleic acid analogs

The present invention provides 5′-modified bicyclic nucleoside analogs and oligomeric compounds comprising at least one of these nucleoside analogs. In preferred embodiments the nucleoside analogs have either (R) or (S)-chirality at the 5′-carbon. These bicyclic nucleoside analogs are useful for enhancing properties of oligomeric compounds including for example enhanced nuclease resistance.

Method of immobilizing and stretching a nucleic acid on a substrate

The present invention relates to a method of immobilizing and stretching a nucleic acid on a silicon substrate, to nucleic acids and substrates prepared according to this method, to uses of the method and to uses of the nucleic acid and the substrate.

Methods of creating and screening dna-encoded libraries

The present invention features a number of methods for identifying one or more compounds that bind to a biological target. The methods include synthesizing a library of compounds, wherein the compounds contain a functional moiety having one or more diversity positions. The functional moiety of the compounds is operatively linked to an initiator oligonucleotide that identifies the structure of the functional moiety.

Methods of using oligomeric compounds comprising 2'-substituted nucleosides

The present disclosure provides oligomeric compounds comprising at least one 2′-fluoroethoxy modified nucleoside of formula I and methods of using these oligomeric compounds. The methods provided herein include contacting a cell or administering to an animal at least one of the oligomeric compounds. In certain embodiments, the oligomeric compounds hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA.

Derivatization of biomolecules by covalent coupling of non-cofactor compounds using methyltransferases

The present invention relates to a use of non-cofactor compounds, represented by formulas (I) or (II) wherein R and Z are independently selected from H, D, C 1 -C 12 -alkyl, preferably C 1 -C 4 -alkyl, alkenyl, alkinyl, phenyl or -LX, wherein X represents a functional group or a reporter group attached via a linker group L, and QH is selected from —SH, —SeH, —NHNH 2 or —ONH 2 , for a targeted modification or derivatization of a biomolecule by covalent coupling to the biomolecule in the presence of a directing methyltransferase. Further development of the method of targeted modification and derivatization are the method for targeted labeling a biomolecule and method for detecting unmethylated target sites in a biomolecule comprising modification of the biomolecule according to the present invention.

Labelling Strategies for the Sensitive Detection of Analytes

The present invention relates to methods and reagents for detecting analytes, e.g. nucleic acids. The new methods and reagents allow a simple and sensitive detection even in complex biological samples.

Flow Chart Programming Platform for Testers and Simulators

A system for the development, compilation, execution, monitoring and debug of automated test and simulation systems in a flow chart programming language. A development and debug system, implemented as software on a computer, which provides an application developer the capability to enter fully defined application programs through the use of graphical flow charts. An executions system, implemented as a program on a device incorporating a central processing unit, memory, communications and necessary interfaces, which executes graphical flow charts compiled by the development and debug system. The development and debug system communicates with the execution system to download programs, control operation, monitor operation and provide a debugging environment.

Preparation and isolation of 5' capped mrna

The synthesis of capped/tagged RNA, methods of use and kits providing same are contemplated. Tagged RNA permits isolation of RNA transcripts in vitro. The ability to isolate and purify capped RNA results in improved transcription and translation and provides a tool for identifying RNA-protein interactions. Such capped RNA finds use in therapeutic applications, diagnosis and prognosis and in the treatment of cancers and HIV.

Bridged artificial nucleoside and nucleotide

It is an object of the present invention to provide a novel molecule for antisense therapies which is not susceptible to nuclease degradation in vivo and has a high binding affinity and specificity for the target mRNAs and which can efficiently regulate expression of specific genes. The novel artificial nucleoside of the present invention has an amide bond introduced into a bridge structure of 2′,4′-BNA/LNA. The oligonucleotide containing the 2′,4′-bridged artificial nucleotide has a binding affinity for a single-stranded RNA comparable to known 2′,4′-BNA/LNA and has an increased nuclease resistance over LNA. Particularly, it is expected to be applied to nucleic acid drugs because of its much stronger binding affinity for single-stranded RNAs than S-oligo's affinity

Modulation of factor 7 expression

Disclosed herein are antisense compounds and methods for decreasing Factor 7 and treating or preventing thromboembolic complications in an individual in need thereof. Examples of disease conditions that can be ameliorated with the administration of antisense compounds targeted to Factor 7 include thrombosis, embolism, and thromboembolism, such as, deep vein thrombosis, pulmonary embolism, myocardial infarction, and stroke. Antisense compounds targeting Factor 7 can also be used as a prophylactic treatment to prevent individuals at risk for thrombosis and embolism.

Modified L-Nucleic Acid

A modified L-nucleic acid, containing an L-nucleic acid part conjugated to a non-L-nucleic acid part is described. The conjugate has extended retention time in and demonstrates a delayed elimination from an organism.

Process for triphosphate oligonucleotide synthesis

The present invention describes simple, efficient, and enzyme-free method of making oligonucleotides with 5′-triphosphate. This invention presents novel process for synthesizing triphosphate oligonucleotides using a diaryl phosphonate as reagent. The process of the present invention is amenable to large-scale, economic 5′-triphosphate oligonucleotide synthesis.

Reversible platelet inhibition

The present invention relates, in general, to receptors and to platelet aggregation and, in particular, to a method of inhibiting platelet aggregation using an aptamer that binds to and inhibits the activity of a receptor, such as glycoprotein IIb/IIIa (gpIIb/IIIa), and to aptamers suitable for use in such a method. The invention also relates to antidotes to antiplatelet agents and to methods of using such antidotes to reverse aptamer-induced platelet inhibition. The invention further relates to von Willebrand Factor (VWF) inhibitors, and antidotes therefore, and to methods of using same.

Modulation of signal transducer and activator of transcription 3 (stat3)expression

Disclosed herein are antisense compounds and methods for decreasing STAT3 mRNA and protein expression. Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate hyperproliferative diseases.

Novel nucleic acid prodrugs and methods of use thereof

Described herein are nucleic acid prodrugs and nucleic acid prodrugs comprising chiral phosphorous moieties. Also described herein are methods of making and using nucleic acid prodrugs and nucleic acid prodrugs comprising chiral phosphorous moieties.

Synthesis of oligonucleotides

A method for preparing an oligonucleotide comprising the steps of a) providing a hydroxyl containing compound having the formula (A), b) reacting said compound with a phosphitylating agent in the presence of an activator (activator I) having the formula (I) to prepare a phosphitylated compound; and c) reacting said phosphitylated compound without isolation with a second compound having the formula (A) wherein R 5 , R 3 , R 2 , B are independently selected, but have the same definition as above in the presence of an activator II different from activator I.

Benzocyanine compounds

Compounds useful as labels with properties comparable to known fluorescent compounds. The compounds are conjugated to proteins and nucleic acids for biological imaging and analysis. Synthesis of the compounds, formation and use of the conjugated compounds, and specific non-limiting examples of each are provided.

In vivo gene regulation by the combination of knock-in-teto sequence into the genome and tetracycline-controlled trans-suppressor (tts) protein

Disclosed is a FAST (Flexible Accelerated STOP TetO-knockin) system, an efficient method for manipulating gene expression in vivo to rapidly screen animal models of disease. This invention further discloses a single gene targeting event yielding 2 distinct knockin mice—STOP-tetO and tetO knockin—which permit generation of multiple strains with variable expression patterns: 1) knockout, 2) Cre-mediated rescue; 3) tTA-mediated misexpression; 4) tTA-mediated overexpression; and 5) tTS-mediated conditional knockout/knockdown. Using the FAST system, multiple gain- and loss-of-function strains can therefore be generated on a timescale not previously achievable. These strains can then be screened for clinically-relevant abnormalities. The flexibility and broad applicability of the FAST system is demonstrated by targeting several genes encoding proteins implicated in neuropsychiatric disorders: Mlc1, Neuroligin 3, the serotonin 1A receptor, and the serotonin 1B receptor.

MODIFIED NUCLEOSIDES, ANALOGS THEREOF AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

The present invention provides modified nucleosides, analogs thereof and oligomeric compounds prepared therefrom. More particularly, the present invention provides modified nucleosides and analogs thereof that are useful for incorporation at the terminus of an oligomeric compound. Such oligomeric compounds can also be included in a double stranded composition. In some embodiments, the oligomeric compounds provided herein are expected to hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. 158.-. (canceled)60. The compound of wherein Mis O.61. The compound of wherein J claim 60 , J claim 60 , Jand Jare each H.62. The compound of wherein Jforms a bridge with one of Jor J.64. The compound of wherein Qand Qare each H.66. The compound of wherein G is halogen claim 65 , OCH claim 65 , OCHF claim 65 , OCHF claim 65 , OCF claim 65 , OCHCH claim 65 , O(CH)F claim 65 , OCHCHF claim 65 , OCHCF claim 65 , OCH—CH═CH claim 65 , O(CH)—OCH claim 65 , O(CH)—SCH claim 65 , O(CH)—OCF claim 65 , O(CH)—N(R)(R) claim 65 , O(CH)—ONO(R)(R) claim 65 , O(CH)—O(CH)—N(R)(R) claim 65 , OCHC(═O)—N(R)(R) claim 65 , OCHC(═O)—N(R)—(CH)—NR)(R) or O(CH)—N(R)—C(═NR)[N(R)(R)] wherein R claim 65 , R claim 65 , Rand Rare each claim 65 , independently claim 65 , H or C-Calkyl.67. The compound of wherein said heterocyclic base moiety is a pyrimidine claim 66 , substituted pyrimidine claim 66 , purine or substituted purine.68. The compound of wherein said heterocyclic base moiety is uracil claim 67 , thymine claim 67 , cytosine claim 67 , 5-methylcytosine claim 67 , adenine or guanine.70. The oligomeric compound of wherein Mis O.71. The oligomeric compound of wherein J claim 70 , J claim 70 , Jand Jare each H.72. The oligomeric compound of wherein Jforms a bridge with one of Jor J.74. The oligomeric compound of wherein Qand Qare each H.76. The oligomeric compound of wherein Ris O and Rand Rare each claim 75 , independently claim 75 , OCH claim 75 , OCHCHor OCH(CH). ...

Fluorescent dyes

Provided are various compounds comprising the formula Also provided are fluorescent dyes comprising the above compound. Additionally, a fluorescence energy transfer system is provided that comprises the above-described fluorescent dye and a second dye, wherein the second dye is capable of energy transfer with the fluorescent dye. Further provided is a kit for labeling a target molecule, where the kit comprises the above-described fluorescent dye with additional reagents useful for labeling the target molecule. Additionally provided is a target molecule labeled with the above-described fluorescent dye. A method of labeling a target molecule is also provided. The method comprises contacting reactive group Z of the above-described fluorescent dye with the target molecule such that reactive group Z reacts with the target molecule to form a covalent bond between reactive group Z and the target molecule. Also, another method of labeling a target molecule is provided. The method comprises contacting the above-described fluorescent dye, which further comprises a member of a binding pair, with the target molecule, where the target molecule comprises a second member of the binding pair.

MODULATION OF INSULIN LIKE GROWTH FACTOR I RECEPTOR EXPRESSION

The present invention provides compositions and methods for modulating the expression of growth factor gene. In particular, this invention relates to compounds, particularly oligonucleotide compounds, which, in preferred embodiments, hybridize with nucleic acid molecules encoding the Insulin Like Growth Factor I receptor (IGF-I receptor or IGF-IR) and in particular human IGF-IR. Such compounds are exemplified herein to modulate proliferation which is relevant to the treatment of proliferative and inflammatory skin disorders and cancer. It will be understood, however, that the compounds can be used for any other condition in which the IGF-IR is involved including inflammatory condition. 144-. (canceled)45. An antisense oligonucleotide comprising 20 to 80 nucleobases in length targeted to a nucleic acid molecule encoding human insulin-like growth factor receptor (IGF-IR) , wherein said oligonucleotide specifically hybridizes with said nucleic acid molecule and inhibits the expression of IGF-IR and wherein said oligonucleotide comprises a deoxynucleotide region flanked on both the 5′ and the 3′ ends of said deoxynucleotide region with 2′-O-(2-methoxyethyl) nucleotides.46. The oligonucleotide as claimed in claim 45 , wherein the oligonucleotide is 20 to 30 nucleobases in length.47. The oligonucleotide as claimed in claim 45 , wherein the oligonucleotide is 8 to 30 nucleobases in length.48. The oligonucleotide as claimed in claim 45 , wherein said oligonucleotide comprises a ten deoxynucleotide region flanked on both the 5′ and the 3′ ends of said ten deoxynucleotide region with five 2′-O-(2-methoxyethyl) nucleotides.49. The oligonucleotide of which is a DNA oligonucleotide claim 45 , an RNA oligonucleotide or a chimeric oligonucleotide.50. The oligonucleotide of claim 45 , wherein the nucleic acid molecule is an RNA molecule encoding human insulin-like growth factor receptor.51. The oligonucleotide of claim 50 , wherein at least a portion of said oligonucleotide ...

Particle matrix for storage of biomolecules

Matrices for manipulation of biopolymers, including the separation, purification, immobilization and archival storage of biopolymers is disclosed.

Procedure for the specific isolation of total dna content of bacterial germs and a kit for this purpose

Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favourably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO 2 —TiO 2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.

Detection or quantification of desirable target molecules, novel dyes, composite dyes, and oligonucleotides or polynucleotides comprising such dyes

The present invention provides dyes, reactive dyes and labeled reagents that may be used in the detection or quantification of desirable target molecules, such as proteins and nucleic acids. Dyes are provided that may be used free in solution where the binding of the dye to the target molecule provides signal generation. Dyes are also provided that comprise reactive groups that may be used to attach the dyes to probes that will bind to desirable target molecules. The novel dyes of the present invention have been modified by the addition of charged and polar groups to provide beneficial properties. 2. The dye of claim 1 , wherein at least one of R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , R claim 1 , or Rfurther comprises a reactive group.3. The dye of claim 2 , wherein said reactive group comprises a nucleophilic reactive group claim 2 , an electrophilic reactive group claim 2 , a terminal alkene claim 2 , a terminal alkyne claim 2 , a coordinate group or an alkylating agent.4. The dye of claim 3 , wherein said nucleophilic reactive group comprises a thiol claim 3 , amine or hydroxyl group.5. The dye of claim 3 , wherein said electrophilic reactive group comprises an isocyanate claim 3 , isothiocyanate claim 3 , monochlorotriazine claim 3 , dichlorotriazine claim 3 , 4 claim 3 ,6 claim 3 ,-dichloro-1 claim 3 ,3 claim 3 ,5-triazines claim 3 , mono- or di-halogen substituted pyridine claim 3 , mono- or di-halogen substituted diazine claim 3 , maleimide claim 3 , haloacetamide claim 3 , aziridine claim 3 , sulfonyl halide claim 3 , acid halide claim 3 , hydroxysuccinimide ester claim 3 , hydroxysulfosuccinimide ester claim 3 , imido ester claim 3 , hydrazine claim 3 , azidonitrophenol claim 3 , azide claim 3 , 3-(2-pyridyl dithio)-proprionamide claim 3 , glyoxal or aldehyde group.6. The dye of claim 1 , wherein said dye further comprises one or more charged groups or polar groups.7. ...

5' MODIFIED NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

The present invention provides 5′ modified nucleosides and oligomeric compounds prepared therefrom. More particularly, the present invention provides modified nucleosides having at least one 5′-substituent and an optional 2′ substituent, oligomeric compounds comprising at least one of these modified nucleosides and methods of using the oligomeric compounds. In some embodiments, the oligomeric compounds provided herein are expected to hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. 146-. (canceled)48. The 5′ modified nucleoside of wherein Bx is uracil claim 47 , thymine claim 47 , cytosine claim 47 , 5-methylcytosine claim 47 , adenine or guanine.49. The 5′ modified nucleoside of wherein qand qare each claim 48 , independently claim 48 , OCH claim 48 , OCHCHor OC(H)(CH)and qis O.50. The 5′ modified nucleoside of wherein r is 1 claim 49 , Mis H and Mis hydroxyl or r is 0 claim 49 , Mis O(CH)CN and Mis N[CH(CH)].51. The 5′ modified nucleoside of wherein G is halogen claim 50 , OCH claim 50 , OCHF claim 50 , OCHF claim 50 , OCF claim 50 , OCHCH claim 50 , O(CH)F claim 50 , OCHCHF claim 50 , OCHCF claim 50 , OCH—CH═CH claim 50 , O(CH)—OCH claim 50 , O(CH)—SCH claim 50 , O(CH)—OCF claim 50 , O(CH)—N(R)(R) claim 50 , O(CH)—ON(R)(R) claim 50 , O(CH)—O(CH)—N(R)(R) claim 50 , OCHC(═O)—N(R)(R) claim 50 , OCHC(═O)—N(R)—(CH)—N(R)(R) or O(CH)—N(R)—C(═NR)[N(R)(R)] wherein R claim 50 , R claim 50 , Rand Rare each claim 50 , independently claim 50 , H or C-Calkyl.52. The 5′ modified nucleoside of wherein G is F claim 51 , OCH claim 51 , O(CH)—OCH claim 51 , OCHC(═O)—N(H)CHor OCHC(═O)—N(H)—(CH)—N(CH).53. The 5′ modified nucleoside of wherein g is 1.54. The 5′ modified nucleoside of wherein three of q claim 53 , q claim 53 , qand qare each H.55. The 5′ modified nucleoside of wherein one of qand qis H claim 53 , one of qand qis H and the other two of q claim 53 , q claim 53 , qand qare other than H.56. The 5′ modified nucleoside of wherein ...

Photogenerated reagents

This invention describes reagent precursors and methods for chemical and biochemical reactions. These reagent precursors that can be activated in solution upon irradiation to generate reagents required for the subsequent chemical reactions. Specifically, photogenerated reagents (PGR) are useful for controlling parallel combinatorial synthesis and various chemical and biochemical reactions. 164-. (canceled)65. A method for forming a nucleic acid polymer comprising:(a) derivatizing a solid surface with a protected moiety containing at least one protective group;(b) contacting the solid surface with solution comprising at least one photogenerated reagent precursor and at least one co-reagent;(c) irradiating the solution thereby generating a photogenerated reagent in at least a portion of the solution such that the protective group is removed from the protected moiety; and(d) coupling a nucleic acid monomer to the deprotected moiety.661. The method of claim wherein the photogenerated reagent precursor is a photogenerated reagent acid precursor.671. The method of claim wherein the co-reagent is a base.681. The method of claim wherein the protected moiety is a protected nucleotide.691. The method of claim wherein the photogenerated reagent precursor is a photogenerated reagent base precursor.701. The method of claim wherein the co-reagent is an acid.711. The method of claim wherein the protected moiety is a protected nucleic acid.721. The method of claim wherein the co-reagent is a sensitizer or supersensitizer.73. A method of deprotecting protected nucleic acids attached to a solid surface and wherein the nucleic acids have one or more protecting group(s) comprising a) contacting the protecting group moieties with a solution comprising one or more PGR-P(s) and one or more co-reagent(s) and b) irradiating the solution to produce one or more PGR(s) wherein the PGR(s) remove the protecting group(s) from the protected nucleic acids.749. The method of claim wherein the ...

SUBSTITUTED 2'-AMINO AND 2'-THIO-BICYCLIC NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

Provided herein are 2′-amino and 2′-thio bicyclic nucleosides and oligomenc compounds prepared therefrom. The novel bicyclic nucleosides provided herein are expected to be useful for enhancing one or more properties of the oligomeric compounds they are incorporated into such as nuclease resistance. 192-. (canceled)94. The bicyclic nucleoside of wherein Bx is an optionally protected uracil claim 93 , thymine claim 93 , cytosine claim 93 , 5-methylcytosine claim 93 , adenine or guanine.95. The bicyclic nucleoside of wherein Tis 4 claim 93 ,4′-dimethoxytrityl and Tis diisopropylcyanoethoxy phosphoramidite.96. The bicyclic nucleoside of wherein Qand Qare each H.97. The bicyclic nucleoside of wherein one of Qand Qis H and the other of Qand Qis C-Calkyl or substituted C-Calkyl.98. The bicyclic nucleoside of wherein one of Qand Qis CH.99. The bicyclic nucleoside of wherein Gand Gare each H.100. The bicyclic nucleoside of wherein one of Gand Gis H and the other of Gand Gis C-Calkyl or substituted C-Calky.101. The bicyclic nucleoside of wherein at least one of Gand Gis CH.102. The bicycle nucleoside of wherein Z is NR wherein R is H claim 93 , C-Csubstituted C-Calkyl claim 93 , C-Calkoxy or substituted C-Calkoxy.103. The bicyclic nucleoside of wherein R is CHor methoxy.104. The bicyclic nucleoside of wherein Z is S.106. The bicyclic nucleoside of wherein three of Q claim 105 , Q claim 105 , Gand Gare H and the other one of Q claim 105 , Q claim 105 , Gand Gis CH.107. The bicyclic nucleoside of wherein two of Q claim 105 , Q claim 105 , Gand Gare H and the remaining two of Q claim 105 , Q claim 105 , Gand Gare CHwherein the two that are CHare selected from Qand G claim 105 , Qand G claim 105 , Qand G claim 105 , and Qand G.109. The oligomeric compound of wherein Bx is an optionally protected uracil claim 108 , thymine claim 108 , cytosine claim 108 , 5-methylcytosine claim 108 , adenine or guanine for each bicyclic nucleoside of Formula II.110. The oligomeric compound of ...

Rapid nucleic acid purification

Provided is a method for rapid nucleic acid purification, and the method for rapid nucleic acid isolation according to the present invention is very useful in diagnosing causes of disease or detecting a target gene; can be used in molecular diagnosis of causes of disease more rapidly and conveniently, as compared with the existing nucleic acid isolation method requiring complicated and special equipment; does not require skills therefor, thereby allowing an ordinary person to personally conduct isolation of nucleic acid for analyzing causes of disease and further solving the existing inconvenience caused by directly going to the hospitals or health clinical centers; and can analyze causes of disease more promptly.

Methods, Reagents and Kits for Preservation of Nucleic Acids in Biological Samples

Provided is a nucleic acid preservative comprising at least one reducing agent, at least one chaotropic substance, at least one polyamine substance and at least one chelating agent and uses thereof, and a method for the preservation of nucleic acids in a biological sample. Further provided are kits for use in the preservation of nucleic acids in a biological sample, and more particularly, a blood sample. 1. A nucleic acid preservative comprising at least one reducing agent , at least one chaotropic substance , at least one polyamine substance and at least one chelating agent.2. The nucleic acid preservative according to claim 1 , wherein the reducing agent is glutathione and wherein the glutathione is present in an amount from about 10 mM to about 200 mM.3. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is a lithium salt and wherein the lithium salt is present in an amount from about 1 M to about 4 M.4. The nucleic acid preservative according to claim 3 , wherein the lithium salt is LiCl.5. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is a guanidium salt claim 1 , and wherein the guanidium salt is present in an amount from about 0.1 M to about 0.9 M.6. The nucleic acid preservative according to claim 5 , wherein the guanidium salt is guanidine hydrochloride.7. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is urea and wherein the urea is present in an amount from about 2 M to about 12 M.8. The nucleic acid preservative according to claim 1 , wherein the polyamine substance is spermidine and wherein the spermidine is present in an amount from about 10 μM to about 300 μM.9. The nucleic acid preservative according to claim 1 , wherein the polyamine substance is biuret and wherein the biuret is present in an amount of about 10 mM to about 100 mM.10. The nucleic acid preservative according to claim 1 , wherein the chelating agent is EDTA and wherein the ...

Bicyclic nucleosides and oligomeric compounds prepared therefrom

The present invention provides novel 3′,5′-linked bicyclic nucleosides and oligomeric compounds prepared therefrom. The bicyclic nucleosides provided herein are useful for enhancing one or more properties of the oligomeric compounds they are incorporated into such as nuclease resistance.

Mechanically-Activated Cation Channels

Methods of screening for agents that modulate the activity of a mechanically-activated cation channel are provided. Also provided are compositions and methods for ameliorating pain by antagonizing or inhibiting mechanically-activated cation channels. 145-. (canceled)46. A method of ameliorating pain in a subject , the method comprising administering to the subject a) an antibody that antagonizes the activity of a mechanically activated cation channel polypeptide of SEQ ID NO:2 , SEQ ID NO:4 , SEQ ID NO:18 , or SEQ ID NO:20 , or b) an antisense oligonucleotide or small interfering RNA (siRNA) complementary to at least 15 contiguous nucleotides of SEQ ID NOs 1 , 3 , 17 , or 19 wherein the antisense oligonucleotide or siRNA inhibits production of the mechanically activated cation channel polypeptide.471. The method of claim , wherein the polypeptide comprises SEQ ID NO:4 or SEQ ID NO:20.481. The method of claim wherein the polypeptide is expressed in bladder , colon , kidney , lung , or skin.491. The method of claim , wherein the polypeptide is expressed in a dorsal root ganglion neuron.501. The method of claim , wherein the subject is a mammal.511. The method of claim , wherein the subject is a human.521. The method of claim , wherein the pain is selected from the group consisting of acute mechanical pain , chronic mechanical pain , mechanical hyperalgesia , mechanical allodynia , arthritis , inflammation , dental pain , cancer pain , and labor pain.531. The method of claim , wherein the antibody is a monoclonal antibody , a humanized antibody or a chimeric antibody.541. The method of claim , wherein the antisense oligonucleotide or siRNA comprises any one of SEQ ID NOs 5-16.55. An isolated antisense oligonucleotide or small interfering RNA (siRNA) complementary to at least 15 contiguous nucleotides of SEQ ID NOs:1 , 3 , 17 , or 19 and encodes a mechanically-activated cation channel polypeptide , wherein the antisense oligonucleotide or siRNA inhibits production of ...

Novel Synthesis of Nucleoside 5'-Triphosphates and Their Derivatives

Disclosed are compounds of nucleoside 5′-triphosphates of formula (I), or derivatives thereof, or pharmaceutically acceptable salts of said nucleoside 5′-triphosphates or said derivatives, wherein the Base of formula (I) is Adenine (A), Cytosine (C), Guanine (G), Thymine (T), Uracil (U), modified nucleobase or unnatural nucleobase; R 1 is H or OH, R 2 is H or OH, X is independently selected from the group consisting of O, S and Se; and Y is independently selected from the group consisting of O, B (borano, or BH 3 ), S, and Se. Also disclosed are processes of preparing the compounds of formula (I), said process comprising steps according to Scheme A:

Method and Kit for Sequential Isolation of Nucleotide Species From a Sample

The invention provides a process and kit for serial isolation of DNA and RNA from the same sample. First, a siliceous solid support with preferential affinity for DNA over RNA is used to capture DNA in a lysate of a sample. Next, a siliceous solid support with similar affinity for RNA and DNA is used to capture RNA from the same lysate. The respective solid supports are recovered independent of each other, washed, and their bound nucleotide species are eluted. The invention further provides DNA and RNA prepared using the process in a minimal number of steps employing a minimal number of reagents. As the invention yields DNA and RNA of high quality and is amenable to automation, the invention may be used widely in the healthcare and pharmaceutical industries. 1. A method of serial recovery of two different nucleic acid species from a biological sample , comprising:in a first binding step, selectively binding a first nucleic acid species to a first solid phase by contacting the biological sample with the first solid phase that selectively binds the first nucleic acid species;separating the first solid phase with the bound first nucleic acid species from an unbound portion of the biological sample;in a second binding step, binding a second nucleic acid species to a second solid phase, different from the first solid phase, by contacting an unbound portion of the biological sample with the second solid phase that binds the first and second nucleic acid species;enzymatic digestion of the first nucleic acid species bound to the second solid phase; andisolating the second nucleic acid species from the second solid phase.2. The method according to claim 1 , further comprising:in the first binding step, homogenization and enzymatic digestion of the biological sample in a lysis buffer, prior to the selectively binding, thereby generating a lysate comprising the first nucleic acid species.3. The method according to claim 1 , wherein the biological sample is lysed in a lysis ...

MODULATION OF EIF4E EXPRESSION

Oligomeric compounds, compositions and methods are provided for modulating the expression of eIF4E. The antisense compounds may be single- or double-stranded and are targeted to nucleic acid encoding eIF4E. Methods of using these compounds for modulation of eIF4E expression and for diagnosis and treatment of diseases and conditions associated with expression of eIF4E are provided. 1. An antisense oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence at least 95% complementary within the 3′ untranslated region (3′UTR) of a nucleic acid encoding human eIF4E , wherein the antisense oligonucleotide comprises at least one chemically modified sugar moiety , internucleoside linkage , or nucleobase.2. The antisense oligonucleotide of claim 1 , wherein the chemically modified sugar moiety is a 2′-O-(2-methoxyethyl) sugar moiety.3. The antisense oligonucleotide of claim 1 , wherein the chemically modified sugar moiety is a bicyclic sugar moiety.4. The antisense oligonucleotide of claim 3 , wherein the bicyclic sugar moiety comprises a 4′-(CH2)-O-2′ or 4′-(CH2)2-O-2′ group.5. The antisense oligonucleotide of claim 1 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.6. The antisense oligonucleotide of claim 1 , wherein the modified nucleobase is a 5-methylcytosine.7. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is chimeric.8. A pharmaceutical composition comprising the antisense oligonucleotide of claim 1 , and a pharmaceutically or physiologically acceptable carrier claim 1 , diluent claim 1 , or excipient.9. The pharmaceutical composition of claim 8 , which is in a form suitable for parenteral administration.10. The pharmaceutical composition of claim 9 , wherein said parenteral administration is via intravenous injection or infusion.11. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide has a nucleobase sequence at least 95% ...

OLIGONUCLEOTIDE SPECIFIC UPTAKE OF NANOCONJUGATES

The present invention concerns nanoparticles functionalized with an oligonucleotide and a domain for a variety of uses, including but not limited to gene regulation. More specifically, the disclosure provides a nanoparticle that is taken up by a cell at an efficiency different than a nanoparticle functionalized with the same oligonucleotide but does not contain a domain. 1. A nanoparticle functionalized with an oligonucleotide and a domain , the nanoparticle having the property of being taken up by a cell at an efficiency different than a nanoparticle functionalized with the same oligonucleotide but lacking the domain.2. The nanoparticle of wherein the domain is located 5′ to the oligonucleotide.3. The nanoparticle of wherein the domain is located 3′ to the oligonucleotide.4. The nanoparticle of wherein the domain is located at an internal region within the oligonucleotide.5. The nanoparticle of wherein the domain is colinear with the oligonucleotide.6. The nanoparticle of functionalized with a second oligonucleotide and the domain is associated with the second oligonucleotide.7. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising more than one thymidine residue.8. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising two thymidine residues.9. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising two claim 1 , three claim 1 , four claim 1 , five claim 1 , six claim 1 , seven claim 1 , eight claim 1 , nine claim 1 , ten claim 1 , eleven claim 1 , twelve claim 1 , thirteen claim 1 , fourteen claim 1 , fifteen claim 1 , sixteen claim 1 , seventeen claim 1 , eighteen claim 1 , nineteen claim 1 , or twenty thymidine residues.10. The nanoparticle of wherein the domain comprises a phosphate polymer (C3 residue).11. The nanoparticle of wherein the domain comprises two or more phosphate polymers (C3 residues).12. A method of modulating cellular uptake ...

MICROORGANISM NUCLEIC ACID PURIFICATION FROM HOST SAMPLES

The present disclosure provides systems, devices, and methods for purifying microorganism nucleic acid from a host sample, such as a whole blood sample from a human. In certain embodiments, devices and systems with multiple filters are employed and provide for the selective removal of blood cells and host nucleic acids from a sample in order to enrich for microorganism nucleic acid. 1. A system or device for purifying microorganism nucleic acid from a sample comprising:i) a first size-exclusion filter that: excludes passage therethrough of lymphocytes, monocytes, neutrophils, eosinophils, and basophils, and allows passage therethrough of erythrocytes, lymphocytes, and platelets; a) a second size-exclusion filter that: excludes passage therethrough of erythrocytes, lymphocytes, and platelets, and allows passage therethrough of microorganisms smaller than platelets; or', I) a second size-exclusion filter that excludes passage therethrough of erythrocytes and lymphocytes, but allows passage therethrough of platelets and microorganisms smaller than platelets, and', 'II) a third size-exclusion filter that excludes passage therethrough of platelets, but allows passage therethrough of microorganisms smaller than platelets; and, 'b) both of the following], 'ii) eitheriii) a shearing filter that mechanically shears said microorganisms such that microorganism nucleic acid is released.2. The system or device of claim 1 , further comprising iv) an affinity capture filter comprising affinity binding molecules that bind human or animal nucleic acid.3. The system or device of claim 2 , wherein said affinity binding molecules bind human repetitive nucleic acid sequences.4. The system or device of claim 1 , wherein said first size-exclusion filter has a pore size of about 10 μm.5. The system or device of claim 1 , wherein the second size-exclusion filter of (ii) (a) has a pore size of about 2 μm or wherein the second size-exclusion filter of (ii) (b) has a pore size of about 5 μm ...

FLUOROPHORE CHELATED LANTHANIDE LUMINESCENT PROBES WITH IMPROVED QUANTUM EFFICIENCY

The invention relates to novel luminescent compositions of matter containing a fluorophore, synthetic methods for making the compositions, macromolecular conjugates of the compositions, and the use of the compositions in various methods of detection. The invention also provides kits containing the compositions and their conjugates for use in the methods of detection. 161.-. (canceled)63. The composition of claim 62 , wherein Ris selected from the group consisting of CF claim 62 , CH claim 62 , and O—CHCH.64. The composition of claim 62 , wherein Ris selected from the group consisting of O—CHCHand O—(CH)—N.65. The composition of claim 62 , wherein Ris selected from the group consisting of H claim 62 , CHC(O)NH—(CH)—N═C═S claim 62 , and CHC(O)NH—(CH)—NHC(O)CHBr.69. The composition of claim 62 , wherein Ris selected from the group consisting of CH claim 62 , CF claim 62 , CHC(O)NH—(CH)—N═C═S claim 62 , and CHC(O)NH—(CH)—NHC(O)CHBr.70. The composition of claim 62 , wherein Ris selected from the group consisting of H and CHC(O)NH—(CH)—N═C═S claim 62 , and CHC(O)NH—(CH)—NHC(O)CHBr.74. The luminescent probe composition of claim 62 , wherein the chelating moiety is selected from the group consisting of EDTA claim 62 , DTPA claim 62 , TTHA claim 62 , DOTA claim 62 , TAGA claim 62 , DOTP claim 62 , DTPA-BMA claim 62 , DO2P claim 62 , and HP-DO3A.75. The composition of further comprising a metal ion chelated to the chelating moiety claim 62 ,wherein the metal is a lanthanide selected from the group consisting of: Gd(III), Dy(III), Ho(III), Er(III), Eu(III), Tb(III), Sm(III), Ce(III), Pr(III), Yb(III), Tm(III), Nd(III), and Tb(IV).76. The composition of claim 62 , wherein the composition is conjugated to a macromolecule claim 62 ,wherein the macromolecule is selected from the group consisting of a polypeptide and a peptide aptamer with binding affinity to a predetermined peptide target.77. The composition of claim 76 , wherein the macromolecule is a polypeptide selected from ...

ANTISENSE NUCLEIC ACIDS

The present invention provides a pharmaceutical composition which causes skipping of the 53rd exon in the human dystrophin gene with a high efficiency. 1. An antisense oligomer which causes skipping of the 53rd exon in the human dystrophin gene , consisting of a nucleotide sequence complementary to any one of the sequences consisting of the 32nd to the 56th or the 36th to the 56th nucleotides from the 5′ end of the 53rd exon in the human dystrophin gene.2. The antisense oligomer according to claim 1 , which is an oligonucleotide.3. The antisense oligomer according to claim 2 , wherein the sugar moiety and/or the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is modified.4. The antisense oligomer according to claim 3 , wherein the sugar moiety of at least one nucleotide constituting the oligonucleotide is a ribose in which the 2′-OH group is replaced by any one selected from the group consisting of OR claim 3 , R claim 3 , R′OR claim 3 , SH claim 3 , SR claim 3 , NH claim 3 , NHR claim 3 , NR claim 3 , N claim 3 , CN claim 3 , F claim 3 , Cl claim 3 , Br and I (wherein R is an alkyl or an aryl and R′ is an alkylene).5. The antisense oligomer according to claim 3 , wherein the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is any one selected from the group consisting of a phosphorothioate bond claim 3 , a phosphorodithioate bond claim 3 , an alkylphosphonate bond claim 3 , a phosphoramidate bond and a boranophosphate bond.6. The antisense oligomer according to claim 1 , which is a morpholino oligomer.7. The antisense oligomer according to claim 6 , which is a phosphorodiamidate morpholino oligomer.911-. (canceled)12. The antisense oligomer according to claim 1 , consisting of the nucleotide sequence shown by SEQ ID NO: 11 or 35.13. A pharmaceutical composition for the treatment of muscular dystrophy claim 1 , comprising as an active ingredient the antisense oligomer according to claim 1 , or ...

PURIFICATION OF SYNTHETIC OLIGONUCLEOTIDES

This invention provides a method for purifying synthetic oligonucleotides comprising capping, polymerizing and separating any failure sequences produced during oligonucleotide synthesis. The invention also provides a method for purifying synthetic oligonucleotides comprising reacting a full length oligonucleotide with a compound to attach a polymerizable functional group to an end of the full length oligonucleotide, polymerizing the full length oligonucleotides and removing the failure sequences, and recovering the full length oligonucleotides. The invention also provides novel capping agents having a polymerizable functional group, and kits comprising at least one composition of the present invention. 2. The method of claim 1 , wherein Ris hydrogen.5. The method of claim 4 , wherein Ar is 1 claim 4 ,4-phenylenyl.6. The method of claim 4 , wherein Y is —O—.7. The method of claim 1 , wherein the failure sequences are polymerized via radical acrylamide polymerization.8. The method of claim 1 , wherein linker is (CH).9. The method of claim 1 , wherein Ris —N(CH(CH)).10. The method of claim 1 , wherein Ris —CH—-CH—CN.11. The method of claim 1 , wherein Ris methyl.12. The method of claim 1 , wherein Rand Rare each hydrogen.14. The method of claim 13 , wherein Rand Rare each hydrogen.17. The method of claim 16 , wherein Ris hydrogen.18. The method of claim 16 , wherein Ar is 1 claim 16 ,4-phenylenyl.19. The method of claim 16 , wherein Y is —O—.20. The method of claim 13 , wherein linker is (CH).22. The method of claim 21 , wherein Ris hydrogen.25. The method of claim 24 , wherein Ar is 1 claim 24 ,4-phenylenyl.26. The method of wherein Y is —O—.28. The method of claim 27 , wherein Ris hydrogen.29. The method of claim 27 , wherein Y is —O—.32. The method of claim 31 , wherein Ar is 1 claim 31 ,4-phenylenyl.33. The method of claim 31 , wherein Y is —O—.34. The method of claim 31 , wherein Z is —O—.36. The method of claim 13 , wherein Ris —N(CH(CH)).37. The method of claim ...

OLIGONUCLEOTIDE AND USE THEREOF

Provided is an oligonucleotide containing an azobenzene derivative, represented by Formula (1) or (2) below: 2. The oligonucleotide according to claim 1 , wherein Rto Rand Rto Rare hydrogen atoms.3. The oligonucleotide according to claim 1 , wherein Rand Rare methyl groups.5. The oligonucleotide according to claim 4 , wherein Rand Rbind with each other to represent a cyclohexyl group or adamantyl group together with a carbon atom for linking to a sulfur atom.6. The oligonucleotide according to claim 4 , wherein{'sup': 31', '32, 'sub': '1-4', 'Rand Reach independently represent a Calkyl group, and'}{'sup': 33', '40, 'Rto Rare hydrogen atoms.'}7. The oligonucleotide according to claim 6 , wherein Rand Rare methyl groups.8. A photo-switching agent claim 1 , by which the formation and dissociation of a double strand can be controlled by visible light irradiation claim 1 , and which is provided with the oligonucleotide according to .10. The photo-switching agent according to claim 9 , wherein Rand Rrepresent methyl groups or hydrogen atoms claim 9 , and R claim 9 , R claim 9 , and Rto Rare hydrogen atoms.11. The photo-switching agent according to claim 9 , wherein each of the pair of oligonucleotides has two or more of the azobenzene derivative adjacent to one another on either side of two or more nucleotides. The present invention relates to an oligonucleotide, and to a photo-switching agent using the oligonucleotide. The priority claim for the present application is based on Japanese Patent Application No. 2010-194942, submitted on Aug. 31, 2010, and the entire contents of the Description of that Japanese Patent Application are incorporated by reference in this Description.Techniques have been developed for controlling hybridization between oligonucleotides with complementary structures by an external stimulus. If hybridization control could be achieved, it could contribute to more precise gene detection, identification and assay, and to the development of molecular ...

Use of 2',5'-oligoadenylate derivative compounds

The invention relates to the therapeutic uses of 2′,5′-oligoadenylate derivative compounds, more particularly for the treatment of chronic fatigue syndrome (CFS) and in the treatment of infection by a gamma-retrovirus.

BUILDING BLOCKS AND METHODS FOR THE SYNTHESIS OF 5-HYDROXYMETHYLCYTOSINE-CONTAINING NUCLEIC ACIDS

The present invention relates to building blocks and methods for the efficient synthesis of 5-hydroxymethylcytosine-containing nucleic acids such as DNA or RNA. 2. The compound of claim 1 , wherein Z is a 5- or 6-membered cyclic radical claim 1 , particularly a ribose claim 1 , ribose analogue or deoxyribose radical claim 1 , wherein the 3′-OH group of the ribose claim 1 , ribose analogue or deoxyribose radical may be substituted by a phosphor-containing group claim 1 , e.g. a phosphate claim 1 , phosphoester or phosphoramidite group and wherein the 5′-OH group of the ribose claim 1 , ribose analogue or deoxyribose radical may be substituted by a protection group.4. The compound of claim 3 , wherein Ris an aliphatic linear or cyclic group comprising up to 6 C-atoms and optionally up to 2 heteroatoms such as N or O claim 3 , e.g. a C(halo) alkyl group claim 3 , or a C(hetero) alkyl group claim 3 , or a Caryl or heteroaryl group optionally substituted by OH claim 3 , halo claim 3 , CN claim 3 , (O)C(halo) alkyl or N(R) claim 3 , wherein Ris as defined for the compound of formula (II).5. A method of introducing a formyl substituent at position 5 of a cytosine cytidine claim 3 , or deoxycytidine compound comprising reacting a 5-halo substituted starting compound with CO under catalysis of Pd.6. A method for the synthesis of a nucleic acid claim 1 , comprising incorporating a compound of into said nucleic acid.7. A method of claim 6 , wherein the nucleic acid synthesis is carried out by a phosphoramidite procedure.8. A nucleic acid molecule having incorporated at least one compound of . The present invention relates to building blocks and methods for the efficient synthesis of 5-hydroxymethylcytosine-containing nucleic acids such as DNA or RNA.The genetic material is constructed from the four canonical bases dA, dC, dG, and dT. The dC base is furthermore subject to epigenetic modification. In eucaryotes the dC base is often methylated at position C5 to give the base 5- ...

Oligonucleotide Conjugates

Oligonucleotide conjugates, where an oligonucleotide is covalently attached to an aromatic system, are provided. In particular embodiments the oligonucleotide is complementary to the RNA component of human telomerase and is covalently attached to a nucleobase via an optional linker. The conjugates inhibit telomerase enzyme activity.

Conformationally restricted dinucleotide monomers and oligonucleotides

This invention relates to conformationally locked dinucleotide motifs for exo- and phosphate stabilization. For instance, oligonucleotides can be prepared having one or more of the following formulas (IV-IX).

LINKERS AND CO-COUPLING AGENTS FOR OPTIMIZATION OF OLIGONUCLEOTIDE SYNTHESIS AND PURIFICATION ON SOLID SUPPORTS

A method of modulation of synthesis capacity on and cleavage properties of synthetic oligomers from solid support is described. The method utilizes linker molecules attached to a solid surface and co-coupling agents that have similar reactivities to the coupling compounds with the surface functional groups. The preferred linker molecules provide an increased density of polymers and more resistance to cleavage from the support surface. The method is particularly useful for synthesis of oligonucleotides, oligonucleotides microarrays, peptides, and peptide microarrays. The stable linkers are also coupled to anchor molecules for synthesis of DNA oligonucleotides using on support purification, eliminating time-consuming chromatography and metal cation presence. Oligonucleotides thus obtained can be directly used for mass analysis, DNA amplification and ligation, hybridization, and many other applications. 1. A compound of the structure:{'br': None, 'sub': s', 'p, 'R-L-R'}{'sub': s', 'p, 'wherein Ris a substrate attaching group, Ris a polymer attaching group, and L is a linker.'}2. The compound of claim 1 , wherein said substrate attaching group is selected from the group consisting of chlorosilyl and alkyloxysilyl functional groups.3. The compound of claim 1 , wherein said polymer attaching group is selected from the group consisting of amine claim 1 , hydroxyl claim 1 , thiol claim 1 , carboxylic acid claim 1 , ester claim 1 , amide claim 1 , epoxide claim 1 , isocyanate claim 1 , and isothiocyanate.4. A method for controlling the number of oligonucleotides synthesized at a predetermined site on a substrate comprising: i) a substrate;', 'ii) a plurality of non-cleavable linkers;', {'sub': '2', 'iii) a plurality of anchor moieties, wherein said anchor moieties comprise the structure —C(X)—C(Y), wherein X comprises —OPOO—, Y is a nucleophile and wherein said structure is part of a ring moiety;'}, 'iv) nucleotide monomers; and', 'v) co-coupling agents;, 'a) providingb) ...

DNA SEQUENCING BY SYNTHESIS USING MODIFIED NUCLEOTIDES AND NANOPORE DETECTION

This disclosure is related to a method of sequencing a single-stranded DNA using deoxynucleotide polyphosphate analogues and translocation of tags from incorporated deoxynucleotide polyphosphate analogues through a nanopore. 5. The method of claim 1 , wherein the tag is ethylene glycol claim 1 , an amino acid claim 1 , a carbohydrate claim 1 , a dye claim 1 , a mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide claim 1 , a fluorescent dyes claim 1 , a chemiluminiscent compound claim 1 , an amino acid claim 1 , a peptide claim 1 , a carbohydrate claim 1 , a nucleotide monophopshate claim 1 , a nucleotide diphosphate claim 1 , an aliphatic acid or an aromatic acid or an alcohol or a thiol with unsubstituted or substituted with one or more halogens claim 1 , a cyano group claim 1 , a nitro group claim 1 , an alkyl group claim 1 , an alkenyl group claim 1 , an alkynyl group claim 1 , an azido group.6. The method of claim 1 , wherein the base is selected from the group consisting of adenine claim 1 , guanine claim 1 , cytosine claim 1 , thymine claim 1 , 7-deazaguanine claim 1 , 7-deazaadenine or 5-methylcytosine.7. The method of claim 1 , further comprising a washing step after each iteration of step (b) to remove unincoporated dNPP analogues from contact with the single-stranded DNA.8. (canceled)9. The method of claim 1 , wherein the single-stranded DNA claim 1 , electrolyte solution and nanopore in the membrane are located within a single container.1011-. (canceled)13. The method of claim 1 , wherein the tag is a mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide and wherein the base of the mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide is the same type of base as the base of the dNPP analogue. ...

OLIGONUCLEOTIDE WITH PROTECTED BASE

The present invention provides a protected nucleotide for elongation, which can be purified efficiently and in a high yield by a liquid-liquid extraction operation, and can achieve an oligonucleotide production method by a phosphoramidite method. 2. The oligonucleotide comprising a protected base of according to claim 1 , wherein q is 0.4. The oligonucleotide comprising a protected base according to claim 3 , wherein R claim 3 , R claim 3 , Rin the number of l claim 3 , Rand Rare each independently a branched chain alkyl group or branched chain alkenyl group selected from the group consisting of a 2 claim 3 ,6 claim 3 ,10 claim 3 ,14-tetramethylpentadecyl group claim 3 , a 2 claim 3 ,6 claim 3 ,10-trimethylundecyl group claim 3 , a 2 claim 3 ,2 claim 3 ,4 claim 3 ,8 claim 3 ,10 claim 3 ,10-hexamethyl-5-undecyl group claim 3 , a 2 claim 3 ,6 claim 3 ,10-trimethylundeca-1 claim 3 ,5 claim 3 ,9-trienyl group claim 3 , a 2 claim 3 ,6-dimethylheptyl group claim 3 , a 2 claim 3 ,6-dimethylhept-5-enyl group claim 3 , a 2 claim 3 ,6-dimethylhepta-1 claim 3 ,5-dienyl group claim 3 , a 9-nonadecyl group claim 3 , a 12-methyltridecyl group claim 3 , an 11-methyltridecyl group claim 3 , an 11-methyldodecyl group claim 3 , a 10-methylundecyl group claim 3 , an 8-heptadecyl group claim 3 , a 7-pentadecyl group claim 3 , a 7-methyloctyl group claim 3 , a 3-methyloctyl group claim 3 , a 3 claim 3 ,7-dimethyloctyl group claim 3 , a 3-methylheptyl group claim 3 , a 3-ethylheptyl group claim 3 , a 5-undecyl group claim 3 , a 2-heptyl group claim 3 , a 2-methyl-2-hexyl group claim 3 , a 2-hexyl group claim 3 , a 3-heptyl group claim 3 , a 4-heptyl group claim 3 , a 4-methyl-pentyl group claim 3 , a 3-methyl-pentyl group claim 3 , and a 2 claim 3 ,4 claim 3 ,4-trimethylpentyl group; or a straight chain alkyl group selected from the group consisting of a tetradecyl group claim 3 , a tridecyl group claim 3 , a dodecyl group claim 3 , an undecyl group claim 3 , a decyl group claim 3 , a ...

Synthesis of oligonucleotides

A method for preparing an oligonucleotide comprising the steps of synthesizing a phosphoramidite by reacting a hydroxyl-containing compound of formula (A) with a phosphitylating agent in the presence of an activator compound of formula (I), to prepare a phosphitylated compound, then coupling the phosphitylated compound without isolation with a second compound having the formula (A), wherein R, R, R, B are independently selected, but have the same definition as above in the presence of an activator II selected from the group of imidazole, imidazolium salts, and mixtures thereof, which are improved activators over activators disclosed in related art. 4. The method of claim 1 , wherein the phosphitylating agent is 2-cyanoethyl-N claim 1 ,N claim 1 ,N′ claim 1 ,N′-tetraisopropylphosphorodiamidite.5. The method of claim 1 , wherein the deprotonated acid is selected from the group consisting of trifluoroacetic acid claim 1 , dichloroacetic acid claim 1 , methanesulfonic acid claim 1 , trifluoromethanesulfonic acid claim 1 , and o-chlorophenolic acid.6. The method of claim 1 , wherein the reacting is in the presence of acetone.7. The method of claim 1 , wherein the concentration of phosphitylating agent in step b) is from 1.0 to 1.2 mol/mol of hydroxyl groups in the hydroxyl containing compound.8. The method of claim 1 , wherein the concentration of phosphitylating agent in step b) is from 3 to 5 mol/mol of hydroxyl groups in the hydroxyl containing compound.9. The method of claim 1 , further comprising adding a polymeric alcohol after step b).10. The method of claim 9 , wherein the polymeric alcohol is polyvinyl alcohol.11. The method of claim 1 , wherein the deprotonated acid is selected from the group consisting of trifluoroacetic acid claim 1 , dichloroacetic acid claim 1 , methanesulfonic acid claim 1 , trifluormethanesulfonic acid (triflate) claim 1 , o-chlorophenolate claim 1 , and mixtures thereof.12. The method of claim 9 , wherein the reacting is in the presence ...

METHODS FOR TREATING MULTIPLE SCLEROSIS USING ANTISENSE OLIGONUCLEOTIDES

A method for treating a patient suffering from multiple sclerosis, particularly a relapsing form of multiple sclerosis, comprising periodically administering a pharmaceutical composition comprising a therapeutically effective amount of OLIGONUCLEOTIDE 1 to the patient, thereby treating the patient. 1. A method for decreasing or inhibiting the accumulation of new active brain lesions , or decreasing or inhibiting an increase in the volume of gadolinium-enhancing brain lesions in a human subject afflicted with multiple sclerosis , comprising periodically administering to the human subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of an oligonucleotide having the structure:{'br': None, 'sup': Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me, '5′-CUG AGT CTG TTT UCC AUU CU-3′'} a) each of the 19 internucleotide linkages of the oligonucleotide is an O,O-linked phosphorothioate diester;', 'b) the nucleotides at the positions 1 to 3 from the 5′ end are 2′-O-(2-methoxyethyl) modified ribonucleosides;', 'c) the nucleotides at the positions 4 to 12 from the 5′ end are 2′-deoxyribonucleosides;', 'd) the nucleotides at the positions 13 to 20 from the 5′ end are 2′-O-(2-methoxyethyl) modified ribonucleosides; and', {'sup': 'Me', 'e) all cytosines are 5-methylcytosines (C),'}], 'wherein'}or a pharmaceutically acceptable salt of the oligonucleotide, so as to thereby decrease or inhibit the accumulation of new active brain lesions, or decrease or inhibit an increase in the volume of gadolinium-enhancing brain lesions in a human subject afflicted with multiple sclerosis.2. The method of claim 1 , wherein the method inhibits progression of disability in the human subject or reduces relapse rate in the human subject.3. The method of claim 95 , wherein the progression of disability is reduced by 15%-70% as measured by EDSS score relative to the progression of disability in a human subject afflicted with ...

Methods and kits for breaking emulsions

The disclosure relates generally to methods, systems, compositions and kits for breaking a water-in-oil emulsion including one or more biomolecules dispersed in an aqueous phase of the water-in-oil emulsion. In some embodiments, the disclosure relates to obtaining a first emulsion including a continuous hydrophobic fraction and a discontinuous aqueous fraction, the aqueous fraction having one or more biomolecules dispersed therein, breaking the first emulsion by contacting the first emulsion with a breaking solution including a second emulsion, where the second emulsion includes a discontinuous phase of organic extraction solvent dispersed in a continuous aqueous phase, and centrifuging to separate the phases of the resulting mixture. In some embodiments, the disclosure relates generally to methods, kits and systems for extracting biomolecules from a water-in-oil emulsion, including breaking a water-in-oil emulsion comprising a plurality of aqueous droplets in a continuous hydrophobic fraction using a breaking solution to produce a resulting reaction mixture containing one or more biomolecules and manipulating the resulting reaction mixture to form at least two phases, where one of the phases includes an aqueous phase containing the one or more biomolecules.

ALIGNMENT OF NANOMATERIALS AND MICROMATERIALS

The present invention provides a method for preparing a nanoassembly that includes the step of reacting the assembly template with at least one nanomaterial to form the nanoassembly using a bifunctional linker. 146.-. (canceled)47. A method of generating an assembly with a desired linear , two-dimensional or three-dimensional structure , comprising:(a) selecting one or more internal positions of a nucleic acid polymer template for attachment of a particle;(b) predicting a linear, two-dimensional and three-dimensional structure of the nucleic acid polymer template when the particle is attached to the one or more selected internal positions of the nucleic acid polymer; [ a single-stranded molecule or a double-stranded molecule comprising DNA, RNA, PNA, or mixed co-polymers thereof, and', 'one or more modified phosphodiester linkages at the selected one or more internal positions within the single stranded molecule or within one or both strands of the double-stranded molecule, wherein the one or more modified phosphodiester linkages each comprise a reactive substituent,, 'wherein the nucleic acid polymer template comprises, 'and wherein the particle comprises at least one linking reagent comprising a first reactive group and a second reactive group separated by a linker segment, wherein the second reactive group is attached the particle,', 'under conditions that permit the first reactive group to attach to the reactive substituent, and, '(c) reacting the nucleic acid polymer template with the particle,'}(d) coupling the particle to the one or more selected internal positions of the nucleic acid polymer template through the linking reagent, thereby forming the assembly with the desired linear, two-dimensional or three-dimensional structure.48. The method of claim 47 , further comprising synthesizing the nucleic acid polymer template claim 47 , wherein synthesizing comprises introducing the reactive substituents at the selected one or more internal positions.49. The ...

OLIGONUCLEOTIDE REPLACEMENT FOR DI-TAGGED AND DIRECTIONAL LIBRARIES

Transposomes and oligonucleotide replacement methods to make DNA libraries that have distinct 5′ and 3′ tags, and to make directional libraries that are enriched for a desired strand. 1. A method for adding a tag to the double-stranded product of a tagmentation reaction , comprising the steps of:(a) providing a double-stranded target nucleic acid and a transposome having a transposase with two transposon end sequences: a “transferred strand” and a “non-transferred strand”;(b) allowing the transposome to fragment the target nucleic acid, whereby the transferred strand is covalently transferred to a first strand of the fragment, and the non-transferred strand remains hybridized to the transferred strand;(c) removing the non-transferred strand from the transferred strand;(d) providing a replacement oligo that comprises a tag sequence, to hybridize to transferred strand; and(e) ligating the replacement oligo to the second strand of the fragment;thereby generating a tagmentation product having a transferred strand and a replacement oligo.2. The method of claim 1 , wherein one strand of the target nucleic acid is chemically modified.3. The method of claim 2 , wherein the chemical modification is conversion of cytosines to uracils.4. The method of claim 1 , wherein the transposome comprises a Tn5 transposase.5. The method of claim 1 , wherein the transposome comprises a Mu transposase.7. The method of claim 6 , wherein the transferred strand further comprises a tag sequence.9. The method of claim 1 , wherein the non-transferred strand is selected from the group consisting of SEQ ID NO:9 claim 1 , SEQ ID NO:10 claim 1 , SEQ ID NO:11 claim 1 , SEQ ID NO:12 claim 1 , SEQ ID NO:13 claim 1 , SEQ ID NO:14 claim 1 , SEQ ID NO:15 claim 1 , SEQ ID NO:16 claim 1 , SEQ ID NO:17 claim 1 , and SEQ ID NO:18.10. The method of claim 1 , wherein step (e) further comprises an extension step.11. The method of claim 2 , further comprising the step of(f) enriching for a desired strand.12. The ...

NEUTRALIZING MONOCLONAL ANTIBODIES AGAINST THE NOGO-66 RECEPTOR (NgR) AND USES THEREOF

The subject invention relates to isolated proteins, particularly monoclonal antibodies, which bind to the Nogo-66 receptor. Specifically, these antibodies have the ability to inhibit the binding of the natural ligand of the Nogo-66 receptor and neutralize the Nogo-66 receptor. These antibodies or portions thereof of the invention are useful for detecting NgR and for inhibiting NgR activity, for example in a human suffering from a disorder in which NgR or Nogo-66 activity is detrimental.

HYBRID SOLID SUPPORTS USEFUL FOR OLIGONUCLEOTIDE PRODUCTION

A method for preparing a crosslinked polymer coated controlled porosity glass (CPG) particle is provided. The method involves mixing CPG particles in a solution comprising polyvinylbenzylchloride and a first solvent at a temperature below 10° C. A second solvent is added and a crosslinking agent is added to the mixture. The first solvent is removed rapidly within 1½ hours of addition of the crosslinking agent. The crosslinking reaction is permitted to proceed and the mixture is then cooled and treated to remove any remaining solvent. The resulting coated CPG particles are washed and dried. Also provided a polymer coated CPG particles using for loading ligand thereon. 1. A conformal polymer-coated controlled porosity glass (CPG) particle for solid phase chemical synthesis comprising:(a) a controlled porosity glass core having pores with a mean average diameter of 500 Angstroms to 4000 Angstroms (50 nm to 4000 nm); and(b) a conformal cross-linked polymeric coating over said core, wherein said polymer comprises a cross-linking agent and monomeric subunits or a polymerizable monomer selected from the group consisting of a vinylbenzylchloride, an acrylic, a styrene, polymers thereof, and mixtures thereof,wherein the conformal polymeric coating layer is on the surface of the CPG particle and within its pores without changing the shape of the coated CPG particle.2. The conformal polymer-coated CPG particle according to claim 1 , wherein the pores within the conformal coated CPG particle retain an average pore size of at least 90% of the average pore size of the uncoated CPG particles (a) claim 1 , based on dry pore size.3. The conformal polymer-coated CPG particle according to claim 1 , wherein the polymeric coating adheres to the CPG core is the absence of a coupling agent to link the coating to the CPG particle.4. The conformal polymer-coated CPG particle according to claim 1 , wherein the polymeric coating is crosslinked in an amount of about 5% to about 30% crosslinked ...

COMPOUNDS AND METHODS FOR MODULATING ANGIOTENSINOGEN EXPRESSION

Disclosed herein are compositions and compounds comprising modified oligonucleotides for modulating AGT and modulating a RAAS pathway related disease, disorder and/or condition in an individual in need thereof. A RAAS pathway related disease, disorder and/or condition in an individual such as hypertension can be treated, ameliorated, delayed or prevented with the administration of antisense compounds targeted to AGT. 1. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of nucleobases 525 to 560 of SEQ ID NO: 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO: 1.2. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of nucleobases 643-691 of SEQ ID NO: 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO: 1.3. The oligomeric compound of claim 1 , wherein the modified oligonucleotide consists of 15 to 30 linked nucleosides.46-. (canceled)7. An oligomeric compound comprising a modified oligonucleotide comprising 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 19 contiguous nucleobases of the nucleobase sequence of SEQ ID NOs: 46 claim 1 , 53-54 claim 1 , 61 claim 1 , 68 claim 1 , 76 claim 1 , 83 claim 1 , 85 claim 1 , 93 claim 1 , 96-97 claim 1 , 109 claim 1 , 134-135 claim 1 , 137-39 claim 1 , 142 claim 1 , 163-172 claim 1 , 180-184 claim 1 , 186 claim 1 , 189 claim 1 , 234 claim 1 , 236 claim 1 , 238-239 claim 1 , 267 claim 1 , 313 claim 1 , 411 claim 1 , 452 claim 1 , 463-470 claim 1 , 475-478 claim 1 , 480 claim 1 , 500-503 claim 1 , 512 claim 1 , 517-518 claim 1 ...

MODULATION OF FACTOR 11 EXPRESSION

Disclosed herein are antisense compounds and methods for decreasing Factor 11 and treating or preventing thromboembolic complications in an individual in need thereof. Examples of disease conditions that can be ameliorated with the administration of antisense compounds targeted to Factor 11 include thrombosis, embolism, and thromboembolism, such as, deep vein thrombosis, pulmonary embolism, myocardial infarction, and stroke. Antisense compounds targeting Factor 11 can also be used as a prophylactic treatment to prevent individuals at risk for thrombosis and embolism. 154.-. (canceled)55. A compound comprising a modified oligonucleotide , wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides , and wherein at least 12 linked nucleosides of the modified oligonucleotide has a nucleobase sequence of SEQ ID NO: 141.56. The compound of claim 55 , wherein the modified oligonucleotide has a nucleobase sequence that is at least 85% complementary to any of the nucleobase sequences of SEQ ID NO: 1 or SEQ ID NO: 2 when measured across the entire nucleobase sequence of the modified oligonucleotide.57. The compound of claim 55 , wherein the modified oligonucleotide consists of a single stranded modified oligonucleotide.58. The compound of claim 55 , wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.59. The compound of claim 58 , wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.60. The compound of claim 55 , wherein the modified oligonucleotide comprises at least one modified sugar.61. The compound of claim 60 , wherein the at least one modified sugar comprises a 2′-O-methoxyethyl group.62. The compound of claim 60 , wherein the at least one modified sugar comprises a 2′-O—CHgroup.63. The compound of claim 60 , wherein the at least one modified sugar is a bicyclic sugar.64. The compound of claim 63 , wherein the bicyclic sugar comprises a 4′-(CH)—O-2′ bridge.65. ...

TRICYCLIC NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

The present invention provides fluorine substituted tricyclic nucleosides of formula (I) wherein: the substituents are as defined in the claims. The present invention further provides oligomeric compounds prepared therefrom. Incorporation of one or more of the tricyclic nucleosides into an oligomeric compound enhances one or more properties of the oligomeric compound. Such oligomeric compounds can also be included in double stranded compositions. 2. The tricyclic nucleoside of claim 1 , wherein Bx is uracil claim 1 , thymine claim 1 , cytosine claim 1 , 5-methylcytosine claim 1 , adenine or guanine.3. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein Tis hydroxyl or protected hydroxyl claim 1 , and wherein Tis a reactive phosphorus group selected from an H-phosphonate or a phosphoramidite.4. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein Tis 4 claim 1 ,4′-dimethoxytrityl and Tis diisopropylcyanoethoxy phosphoramidite or a controlled pore glass surface.5. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein one or two of q claim 1 , q claim 1 , qzand zis F claim 1 , and the other of q claim 1 , q claim 1 , qzand zare H.8. The oligomeric compound of claims 7 , wherein each Bx is claims 7 , independently claims 7 , uracil claims 7 , thymine claims 7 , cytosine claims 7 , 5-methylcytosine claims 7 , adenine or guanine.9. The oligomeric compound of any one of to claims 7 , wherein each internucleoside linking group is claims 7 , independently claims 7 , a phosphodiester internucleoside linking group or a phosphorothioate internucleoside linking group.10. The oligomeric compound of any one of to claims 7 , wherein essentially each internucleoside linking group is a phosphorothioate internucleoside linking group.11. The oligomeric compound of any one of to claims 7 , comprising a first region having at least two contiguous tricyclic nucleosides having Formula II.12. The oligomeric compound ...

Protected Monomer and Method of Final Deprotection for RNA Synthesis

A method of deprotecting a solid support bound polynucleotide includes the step of contacting the polynucleotide with a composition comprising a diamine under conditions sufficient to deprotect the 2′-protected ribonucleotide residue. The solid support bound polynucleotide has at least one 2′-protected ribonucleotide residue, which has the following structure: wherein B P is a protected or unprotected heterocycle; R 12 is a protecting group selected from a hydrocarbyl, a substituted hydrocarbyl, an aryl, and a substituted aryl; X is O or S; and PG is a thionocarbamate protecting group.

FLUORESCENT DYES AND METHODS OF USE THEREOF

Provided are methods for labeling target molecules, such as nucleic acids, with fluorescent dye compounds having the formula 1. A composition of matter , comprising:a first PCR primer of a PCR primer pair, said first PCR primer comprising a first oligonucleotide covalently labeled with Dye 1 or Dye 7.2. The composition of matter of claim 1 , wherein the first oligonucleotide is covalently labeled with Dye 1.3. The composition of matter of claim 2 , further comprising:a second PCR primer of the PCR primer pair, said second PCR primer comprising a second oligonucleotide.4. The composition of matter of claim 3 , wherein said second oligonucleotide is covalently labeled with a fluorophore.5. The composition of matter of claim 4 , wherein said fluorophore is fluorescein.6. The composition of matter of claim 1 , wherein the first oligonucleotide is covalently labeled with Dye 1.7. The composition of matter of claim 6 , further comprising:a second PCR primer of the PCR primer pair, said second PCR primer comprising a second oligonucleotide.8. The composition of matter of claim 7 , wherein said second oligonucleotide is covalently labeled with a fluorophore.9. The composition of matter of claim 8 , wherein said fluorophore is fluorescein.10. A polynucleic acid molecule comprising nucleotides wherein at one of said nucleotides is labeled with Dye 1 or Dye 7.11. The polynucleic acid molecule of claim 10 , wherein at least one of said nucleotides is labeled with Dye 1.12. The polynucleic acid molecule of claim 10 , wherein at least one of said nucleotides is labeled with Dye 7. This application is a continuation of U.S. application Ser. No. 15/205,438 filed Jul. 8, 2016, which is a divisional of U.S. application Ser. No. 13/270,456 filed Oct. 11, 2011 (now U.S. Pat. No. 9,416,153), each of which is hereby incorporated by reference in its entirety.The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby ...

TRIAZABUTADIENES AS CLEAVABLE CROSS-LINKERS

Triazabutadiene molecules as cleavable cross-linkers adapted to cross-link components with click chemistry, e.g., clickable triazabutadienes. For example, in some embodiments, the triazabutadienes feature alkyne handles attached to the imidazole portion or the aryl portion of the triazabutadienes, wherein the alkyne handles can link to azide handles (e.g., azide handles disposed on other components) via click chemistry. Also described are methods of producing said clickable triazabutadienes and methods of use of said clickable triazabutadienes. The present invention also features methods of cleaving said clickable triazabutadienes, e.g., for liberating the diazonium species for further chemical reactions. 120-. (canceled)22. The clickable triazabutadiene of claim 21 , wherein the tri-substituted aryl group comprises mesityl claim 21 , a NHS-ester moiety; an oligonucleotide; a peptide; a fluorescence quencher; a pro-fluorophore; an alkyne; a triazene; an aldehyde; an amine; an aminooxy; a halogen; or a combination thereof.23. The clickable triazabutadiene of claim 21 , wherein the triazabutadiene comprises is linked to a peptide claim 21 , an oligonucleotide claim 21 , or a drug.24. The clickable triazabutadiene of claim 21 , wherein the linking component comprises a peptide claim 21 , an oligonucleotide claim 21 , or a drug.26. The clickable triazabutadiene of claim 25 , wherein the tri-substituted aryl group comprises mesityl claim 25 , a NHS-ester moiety; an oligonucleotide; a peptide; a fluorescence quencher; a pro-fluorophore; an alkyne; a triazene; an aldehyde; an amine; an aminooxy; a halogen; or a combination thereof.27. The clickable triazabutadiene of claim 25 , wherein the triazabutadiene comprises is linked to a peptide claim 25 , an oligonucleotide claim 25 , or a drug.28. The clickable triazabutadiene of claim 25 , wherein the linking component comprises a peptide claim 25 , an oligonucleotide claim 25 , or a drug. Triazabutadienes can be triggered to ...

Novel Compounds

A compound of Formula (I) 2. The method of wherein the cancer is melanoma.3. The method of wherein the cancer is head and neck cancer.4. The method of wherein the compound is administered by intratumoral injection.5. The method of wherein the compound is administered by peritumoral injection.6. The method of wherein the cancer is squamous cell carcinomas.7. The method of wherein the cancer is breast cancer.8. The method of wherein the compound is administered by intratumoral injection.9. The method of wherein the compound is administered by peritumoral injection.10. The method of wherein the cancer is hepatocellular cancer.11. The method of wherein the cancer is colon cancer.12. The method of wherein the cancer is esophageal cancer.13. The method of wherein the cancer is rectal cancer.14. The method of wherein the cancer is lung cancer.15. The method of wherein the compound is administered by intratumoral injection.16. The method of wherein the compound is administered by peritumoral injection.17. The method of wherein the cancer is renal cell cancer.18. The method of wherein claim 1 , the cancer is selected from: melanoma claim 1 , squamous cell carcinomas claim 1 , hepatocellular cancer claim 1 , colon cancer claim 1 , esophageal cancer claim 1 , rectal cancer claim 1 , and renal cell cancer claim 1 , and the compound is administered by intratumoral injection.19. The method of claim 1 , the wherein claim 1 , the cancer is selected from: melanoma claim 1 , squamous cell carcinomas claim 1 , hepatocellular cancer claim 1 , colon cancer claim 1 , esophageal cancer claim 1 , rectal cancer claim 1 , and renal cell cancer claim 1 , and the compound is administered by peritumoral injection.20. The method of wherein the cancer is selected from: brain (gliomas) claim 1 , glioblastomas claim 1 , astrocytomas claim 1 , glioblastoma multiforme claim 1 , Bannayan-Zonana syndrome claim 1 , Cowden disease claim 1 , Lhermitte-Duclos disease claim 1 , Wilm's tumor claim 1 , Ewing' ...

Phosphorodiamidate Backbone Linkage for Oligonucleotides

This invention relates to antisense oligonucleotides comprising at least one N3′→P5′ phosphorodiamidate linkage (NPN) in the backbone as well as methods for using the same. The antisense oligonucleotides can effectively prevent or decrease protein expression. 114.-. (canceled)16. The compound of claim 15 , or a salt thereof claim 15 , wherein W is O.17. The compound of claim 15 , or a salt thereof claim 15 , wherein Bis independently selected from purinyl and pyrimidinyl.18. The compound of claim 15 , or a salt thereof claim 15 , wherein Bis independently selected from 4-benzoyl-1-cytosinyl claim 15 , 6-benzoyl-9-adeninyl claim 15 , 6-dimethylformamidino-9-adeninyl claim 15 , 2-isobutyryl-9-guaninyl claim 15 , 2-dimethylformamidino-9-guaninyl claim 15 , 9-adeninyl claim 15 , 9-guaninyl claim 15 , 1-cytosinyl claim 15 , 1-thyminyl and 1-uracilyl.19. The compound of claim 15 , or a salt thereof claim 15 , wherein Bis selected from adenine claim 15 , guanine claim 15 , cytosine claim 15 , thymine claim 15 , and uracil.20. The compound of claim 15 , or a salt thereof claim 15 , wherein Ris hydrogen.21. The compound of claim 15 , or a salt thereof claim 15 , wherein Rand Rare methyl.22. The compound of claim 15 , or a salt thereof claim 15 , wherein Ris selected from trityl claim 15 , dimethoxytrityl and methoxytrityl.23. The compound of claim 15 , or a salt thereof claim 15 , wherein Ris selected from chloro claim 15 , iodo claim 15 , bromo claim 15 , fluoro claim 15 , methanesulfonyloxy claim 15 , tosyloxy claim 15 , triflyloxy claim 15 , nitro-phenylsulfonyloxy and bromo-phenylsulfonyloxy.24. The compound of claim 15 , or a salt thereof claim 15 , wherein Ris halogen.25. The compound of claim 15 , or a salt thereof claim 15 , wherein Ris selected from trityl claim 15 , dimethoxytrityl and methoxytrityl and Ris halogen.2640.-. (canceled)4260.-. (canceled)62. The method of claim 61 , wherein the phosphorylating reagent is dimethylaminophosphoryl compound.63. The method ...

Flex plate with removable inserts and cover

Technologies are described for methods and systems effective for flex plates. The flex plates may comprise a base plate. The base plate may include walls that define an insert location opening in the base plate. The insert location opening in the base plate may be in communication with a securement area. The flex plates may comprise an insert. The insert may include a reservoir region and a crystallization region separated by a wall including channels. The reservoir region and the crystallization region may include a backing. The insert may further include securement tabs. The securement tabs may be configured to secure the insert to the base plate at the securement area.

A METHOD FOR GENERATING RANDOM OLIGONUCLEOTIDES AND DETERMINING THEIR SEQUENCE

Random oligonucleotides are generated with incomplete information about the sequence of the nucleic acid bases present in the newly generated molecules. The sequences of the oligonucleotides are subsequently determined and then these oligonucleotides can be processed for various potential uses. 1. A method of generating an oligonucleotide , the method comprising:a. generating at least one molecule comprising nucleotides by adding at least one nucleotide at random to the molecule, wherein the molecule generated is a random oligonucleotide;b. determining the sequence of the random oligonucleotide; andc. selecting random oligonucleotides using certain characteristics of the random oligonucleotides.2. The method of wherein the random oligonucleotides are generated using phosphoramidite chemistry.3. The method of claim 1 , wherein the random oligonucleotides are generated using an enzymatic process.4. The method of claim 1 , wherein the random oligonucleotides are generated within a microwell.5. The method of claim 1 , wherein the random oligonucleotides are generated on a microarray.6. The method of wherein the characteristic used to select the random oligonucleotide is a specific sequence of nucleotides.7. The method of wherein the characteristic to select the random oligonucleotide is a size of the random oligonucleotide.8. The method of wherein the random oligonucleotides are generated on oligonucleotides having a sequence that is at least partially known.9. The method of claim 3 , wherein an indicator molecule becomes reactive after a nucleotide is added to the molecule.10. The method of wherein adding a nucleic acid base to the molecules is partially directed.11. The method of claim 1 , wherein microfluids are used to control reaction conditions.12. The method of wherein directed energy is used to control the reaction conditions.13. The method of where the properties of the random oligonucleotide are measured using a nanopore.14. The method of wherein the selected ...

LIGHT-STIMULATED RELEASE OF CARGO FROM OLIGONUCLEOTIDES

The invention provides oligonucleotide conjugates including a photolabile crosslinker attached to a cargo moiety, e.g., a therapeutic or diagnostic agent. The invention further provides reagents useful in the preparation of such conjugates and methods of their use. 2. The conjugate of claim 1 , wherein L is present and amido claim 1 , C1-10 alkylene claim 1 , or C1-20 polyalkeneoxide.3. The conjugate of claim 1 , wherein L is present and C2-C20 polyethylene glycol.4. The conjugate of claim 1 , wherein Ais triazolyl claim 1 , disulfide claim 1 , cyclohexenyl claim 1 , amido claim 1 , thioamido claim 1 , acetal claim 1 , ketal claim 1 , or sulfonamido.5. The conjugate of claim 1 , wherein Y is C1-10 alkyl.6. The conjugate of claim 1 , wherein Y is methyl.7. The conjugate of claim 1 , wherein Ais the amine reactive group.8. The conjugate of claim 7 , wherein the amine reactive group is p-nitrophenoxyl claim 7 , N-hydroxysuccinimidyl claim 7 , halide claim 7 , pentafluorophenoxyl claim 7 , or imidazolyl.9. The conjugate of claim 1 , wherein Ais —NHX.10. The conjugate of claim 9 , wherein X is a therapeutic or diagnostic agent.13. The crosslinker of claim 12 , wherein L is present and C1-11 alkylene or C1-20 polyalkeneoxide.14. The crosslinker of claim 12 , wherein L is present and C2-C20 polyethylene glycol.15. The crosslinker of claim 12 , wherein Ais azido claim 12 , alkynyl claim 12 , alkenyl claim 12 , thiol claim 12 , halide claim 12 , boronic acid claim 12 , hydroxyl claim 12 , carboxyl claim 12 , formyl claim 12 , or ketone.16. The crosslinker of claim 12 , wherein Y is C1-11 alkyl.17. The crosslinker of claim 12 , wherein Y is methyl.18. The crosslinker of claim 12 , wherein Ais p-nitrophenoxyl claim 12 , N-hydroxysuccinimidyl claim 12 , halide claim 12 , pentafluorophenol claim 12 , or imidazolyl.20. An oligonucleotide conjugate comprising an oligonucleotide of 2-1000 nucleotides in length claim 12 , conjugated to a therapeutic or diagnostic agent by a ...

Oligonucleotide compositions and methods thereof

Among other things, the present disclosure relates to designed oligonucleotides, compositions, and methods thereof. In some embodiments, provided oligonucleotide compositions provide altered splicing of a transcript. In some embodiments, provided oligonucleotide compositions have low toxicity. In some embodiments, provided oligonucleotide compositions provide improved protein binding profiles. In some embodiments, provided oligonucleotide compositions have improved delivery. In some embodiments, provided oligonucleotide compositions have improved uptake. In some embodiments, the present disclosure provides methods for treatment of diseases using provided oligonucleotide compositions.

Cyclic Di-Nucleotide Compounds as STING Agonists

A class of polycyclic compounds of general formula (I), wherein Base1, Base2, Y, Ya, Xa, Xa1, Xb, Xb1, Xc, Xc1, Xd, Xd1, R1, R1a, R2a, R3, R3a, R4, R5, R5a, R6, R6a, R7, R7a, R8, R8a, and R9 are defined herein, that may be useful as inductors of type I interferon production, specifically as STING active agents, are provided. Also provided are processes for the synthesis and use of compounds (I).

Mobile Solid Phase Reaction System and Method

A system and method are disclosed. A system for contacting a mobile solid phase with a flowing fluid phase includes: one or more reaction module, wherein the one or more reaction module comprises a conduit for the passage of a fluid phase and a solid phase, the conduit comprising a fluid input port and a fluid outlet port, and a first service module operably connected to a first side of a reaction module, the first service module for supplying and/or receiving the fluid phase to and/or from the reaction module, wherein the system is configured for passing a solid phase through the reaction module, via the conduit. 1. A system for contacting a mobile solid phase with a flowing fluid phase comprising: 'wherein the first reaction module comprises a conduit for the passage of a fluid phase and a solid phase, the conduit comprising a fluid input port and a fluid outlet port; and', 'a first reaction module;'}a first service module operably connected to a first side of the first reaction module, the first service module for supplying and/or receiving the fluid phase to and/or from the first reaction module;wherein the system is configured for passing a solid phase through the first reaction module, via the conduit.2. The system as claimed in further comprising a second reaction module claim 1 , provided in series claim 1 , such that the solid phase may pass through the consecutive reaction modules.3. The system as claimed in wherein the two reaction modules claim 2 , and the first service module claim 2 , are all configured to releasably connect to adjacent modules.4. The system as claimed in claim 1 , wherein the first side and a further side of the first reaction module are each a mating face; andwherein the first service module has a mating face that is connectable with a respective mating face of the first reaction module.5. The system as claimed in claim 1 , wherein the conduit of the first reaction module comprises a solid phase input port and a solid phase output ...

Locked nucleic acid inhibitor mir-145 and uses thereof

The present invention provides oligonucleotides with chemical motifs that are miR-145 inhibitors. The oligonucleotides can be used for the treatment and prevention of a condition by inhibiting the expression or activity of miR-145 in cells of a subject in need thereof. Methods provided include treating or preventing pulmonary arterial hypertension, neointima formation, restenosis or hypertension in a subject in need thereof by administering to the subject an inhibitor of miR-145 expression or activity. Pharmaceutical compositions and kits comprising miR-145 inhibitors are also disclosed.

DIAMINOPHENOTHIAZINIUM DERIVATIVES FOR LABELLING BIOMOLECULES, METHOD AND SUBSTRATE FOR LABELLING OLIGONUCLEOTIDES, AND OLIGONUCLEOTIDES OBTAINED

The present invention relates to diaminophenothiazinium derivatives of formula (I); in which R, R, R, R, R, Rand X are as defined in Claim and also the methods for labelling oligonucleotides using such a derivative, labeling substrates and the oligonucleotides which can be obtained by means of such methods or from such labelling substrates. 2. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that Aand Aare linear or branched alkylene chains in which from 2 to 6 consecutive carbon atoms separate the oxygen and nitrogen atoms.3. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that at least one of the R claim 1 , R claim 1 , Rand Rgroups does not represent an -A-ORgroup as defined in and said R claim 1 , R claim 1 , Ror Rgroup(s) different than -A-ORand that -A-ORrepresent(s) an alkyl group having from 2 to 12 carbon atoms claim 1 , preferably from 4 to 12 carbon atoms.4. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that R═R═H.5. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that Rrepresents a —P{N[(C-C)alkyl]}(OCHCHC≡N) group claim 1 , such as the —P[N(Pr)](OCHCHC≡N) group.9. A method for labeling an oligonucleotide with a diaminophenothiazinium derivative as claimed in claim 1 , which comprises the growth of an oligonucleotide grafted onto a solid substrate claim 1 , and the replacement of one or more of the nucleotides of which it is formed with one or more of said diaminophenothiazinium derivatives claim 1 , before the oligonucleotide is detached from the solid substrate.10. The labeling method as claimed in claim 9 , characterized in that at least one replacement with a diaminophenothiazinium derivative is carried out before the end of the growth of the oligonucleotide.11. The labeling method as claimed in claim 9 , characterized in that at least one substitution with a diaminophenothiazinium derivative is carried out in the 3′ or 5′ ...

TARGETED TRANSPOSITION FOR USE IN EPIGENETIC STUDIES

Disclosed herein are compositions, methods and kits useful for epigenetic analysis based on the use of transposons that are targeted to specific regions of chromatin based on DNA-DNA interactions, protein-protein interactions, RNA-RNA interactions, and nucleic acid-protein interactions. 1. A method of making a nucleic acid sequence library or libraries comprising:a. extracting chromatin from a sample;b. adding at least one protein-oligonucleotide conjugate comprising an extraction moiety to said chromatin;c. allowing said protein conjugates to locate at its/their target proteins and/or target DNA-binding sites and/or noncoding RNA-binding sites in said chromatind. tagging the nucleic acid in said chromatin fragments with said conjugate by inducing an intermolecular reaction between said oligonucleotide and said nucleic acid; ande. extracting the nucleic acids tagged using the extraction moiety.2. A method according to wherein the protein-oligonucleotide conjugate comprises transposase and the intermolecular reaction is transposition.3. A method of making a nucleic acid sequence library or libraries comprising:a. extracting and optionally fragmenting chromatin from a sample;b. adding at least one oligonucleotide-transposome construct comprising an extraction moiety to said chromatin;c. allowing said oligonucleotide-transposome construct to locate at its/their target DNA and/or noncoding RNA-binding sites in said chromatin;d. tagging the nucleic acid in said chromatin fragments with said construct by inducing an intermolecular reaction between said oligonucleotide and said nucleic acid; ande. extracting the nucleic acids tagged using the extraction moiety.4. A method according to wherein the oligonucleotide-transposome construct comprises transposase and the intermolecular reaction is transposition.5. The oligonucleotide of claim 3 , wherein the oligonucleotide targets non-coding RNA.6. The oligonucleotide of claim 3 , wherein the oligonucleotide contains peptide ...

LABELLED NUCLEOTIDES

The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterised in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH and NQ wherein Q is a Csubstituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH and N(allyl), T is hydrogen or a Csubstituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside). 1. (canceled)2. (canceled)4. The oligonucleotide of claim 3 , wherein R is not present.6. The oligonucleotide of claim 5 , wherein the dotted line connecting the Fluor and the benzene ring comprises one or more spacer units such that the Fluor is held a sufficient distance from the nucleobase so as not to interfere with any interaction between the nucleotide and an enzyme.7. The oligonucleotide of claim 6 , wherein the spacer unit comprises —O— claim 6 , —NH— claim 6 , or —N(alkyl)-.9. The oligonucleotide of claim 8 , wherein the Fluor comprises a cyanine claim 8 , a rhodamine claim 8 , or a coumarin dye.10. The oligonucleotide of claim 9 , wherein the Fluor is a rhodamine dye.11. The oligonucleotide of claim 9 , wherein the Fluor is a Cy5 dye.12. The oligonucleotide of claim 9 , wherein the Fluor is attached to the Linker by reacting an N-hydroxysuccinimide ester of the Fluor with an amino moiety of the Linker.17. The oligonucleotide of claim 8 , wherein the oligonucleotide is in contact with an aqueous solution comprising ethylenediaminetetraacetic acid (EDTA).18. The oligonucleotide of claim 8 , wherein the oligonucleotide is in contact with a mutant polymerase.19. The oligonucleotide of claim 8 , wherein the oligonucleotide is in contact with a water soluble phosphine.20. The oligonucleotide of claim 8 , wherein the oligonucleotide is hybridized to a target polynucleotide claim 8 , and wherein the ...

Labelled nucleotides

The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterised in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH and NQ wherein Q is a C 1-10 substituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH and N(allyl), T is hydrogen or a C 1-10 substituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside).

Cyclic dinucleotides as sting agonists

Disclosed are compounds, compositions and methods for treating of diseases, syndromes, or disorders that are affected by the modulation of STING. Such compounds are represented by Formula (I) as follows: wherein B 2 , X 2 , R 2a , R 2b , R 2c , Z-M-Y, Y 1 -M 1 Z 1 , B 1 , X 1 , R 1a , R 1b , R 1c are as defined herein.

Modified L-Nucleic Acid

A modified L-nucleic acid, containing an L-nucleic acid part conjugated to a non-L-nucleic acid part is described. The conjugate has extended retention time in and demonstrates a delayed elimination from an organism. 1. (canceled)2. A modified L-nucleic acid , comprising a L-nucleic acid part and a non-L-nucleic acid part , wherein the L-nucleic acid part is conjugated with the non-L-nucleic acid part , and the conjugation of the L-nucleic acid part with the non-L-nucleic acid part leads to an increased retention time in an organism or a retarded excretion from an organism compared to a L-nucleic acid comprising only the L-nucleic acid part , and wherein said L-nucleic acid part is a spiegelmer.3. The modified L-nucleic acid of claim 2 , wherein the non-L-nucleic acid part has a molecular weight of more than about 300 Da.4. The modified L-nucleic acid of claim 2 , wherein the modified L-nucleic acid has a molecular weight of about 600 to 500 claim 2 ,000 Da.5. The modified L-nucleic acid of claim 2 , wherein the L-nucleic acid part has a molecular weight of 300 to 50 claim 2 ,000 Da.6. The modified L-nucleic acid of claim 2 , wherein the non-L-nucleic acid part is linked to the L-nucleic acid part via a functional group of the L-nucleic acid part claim 2 , wherein the functional group is selected from the group consisting of terminal and non-terminal phosphates claim 2 , terminal and non-terminal sugar portions claim 2 , natural and non-natural purine bases claim 2 , and natural and non-natural pyrimidine bases.7. The modified L-nucleic acid of claim 6 , wherein the linkage of the non-L-nucleic acid part with the L-nucleic acid part is via the 2′-OH— claim 6 , 3′-OH— claim 6 , 5′-OH-group or a derivative therefrom claim 6 , or one or more sugars of the L-nucleic acid part.8. The modified L-nucleic acid of claim 6 , wherein the linkage is via at least one of the positions 5 or 6 of a pyrimidine base.9. The modified L-nucleic acid of claim 6 , wherein the linkage is ...

Nucleic acid polyhedra from self-assembled vertex-containing fixed-angle nucleic acid structures

Provided herein are compositions comprising nucleic acid structures comprising three or more arms arranged at fixed angles from each other, composites thereof such as DNA cages, and methods for their synthesis and use.

Method for detecting the presence of a target nucleic acid sequence in a sample

A method comprises loading a sample portion into a sample chamber which comprises means for minimizing diffusion of the sample portion, subjecting the sample portion to an amplification step, and determining whether the sample portion contains at least one molecule of a target nucleic acid. If the sample portion contains a single molecule of the target nucleic acid, the sample portion would attain a detectable concentration of the target nucleic acid after a single round of amplification. Also, a microfluidic device comprising a sample portion and a sample chamber comprising means for minimizing diffusion of the sample portion. Also, a microfluidic device comprising a sample chamber and an amplification targeting reagent positioned in the first sample chamber.

SOLID SUPPORTS AND PHOSPHORAMIDITE BUILDING BLOCKS FOR OLIGONUCLEOTIDE CONJUGATES

Novel non-nucleoside solid supports and phosphoramidite building blocks for preparation of synthetic oligonucleotides containing at least one non-nucleosidic moiety conjugated to a ligand of practical interest and synthetic processes for making the same are disclosed. Furthermore, oligomeric compounds are prepared using said solid supports and phosphoramidite building blocks, preferably followed by removal of protecting groups to provide oligonucleotides conjugated to ligands of interest. 2. The compound of wherein one of R and Ris selected from tris-(4-methoxyphenyl)methyl protecting group claim 1 , bis-(4-methoxyphenyl)phenylmethyl protecting group claim 1 , 9-phenylxanthen-9-yl protecting group claim 1 , or 9-(4-methoxyphenyl)xanthen-9-yl protecting group and the other of R and Ris selected from a residue of succinic acid optionally further attached to a solid phase material W via the second carboxylic function or a residue of diglycolic acid optionally further attached to a solid phase material W via the second carboxylic function.3. The compound of wherein one of R and Ris selected from tris-(4-methoxyphenyl)methyl protecting group claim 1 , bis-(4-methoxyphenyl)phenylmethyl protecting group claim 1 , 9-phenylxanthen-9-yl protecting group claim 1 , or 9-(4-methoxyphenyl)xanthen-9-yl protecting group and the other of R and Ris a phosphoramidite moiety PA.4. The compound of wherein each Rand Ris isopropyl group or Rand Rtogether with the nitrogen they are attached to form a cycle so that R+R=—(CH)— claim 3 , R+R=—(CH)— claim 3 , or R+R=—(CH)—O—(CH)—.5. The compound of wherein each A and Ais independently selected from —CH— or —(CH)—.6. The compound of wherein each E and Eis independently selected from —CH— claim 1 , —OCH— claim 1 , —(CH)— claim 1 , or —O(CH)—.7. The compound of wherein G is selected from an atom of hydrogen claim 1 , an alkyl group claim 1 , a trifluoroacetyl group claim 1 , (9H-fluoren-9-yl)methoxycarbonyl (Fmoc) group claim 1 , 6-( ...

Oligonucleotide Compositions and Methods of Making the Same

The present disclosure provides a solid phase method of making oligonucleotides via sequential coupling cycles including at least one coupling of a dinucleotide dimer subunit to a free 3′-terminal group of a growing chain. The oligonucleotides include at least two nucleoside subunits joined by a N3′→P5′ phosphoramidate linkage. The method may include the steps of (a) deprotecting the protected 3′ amino group of a terminal nucleoside attached to a solid phase support, said deprotecting forming a free 3′ amino group; (b) contacting the free 3′ amino group with a 3′-protected amino-dinucleotide-5′-phosphoramidite dimer in the presence of a nucleophilic catalyst to form an internucleoside N3′→P5′ phosphoramidite linkage; and (c) oxidizing (e.g., sulfurizing) the linkage. The compositions produced by the subject methods may include a reduced amount of one or more (N−x) oligonucleotide products. Also provided are pharmaceutical compositions including the subject oligonucleotide compositions. 141.-. (canceled)43. The method of claim 42 , wherein oxidizing the linkage comprises sulfurization to produce a thiophosphoramidate linkage.44. The method of claim 42 , wherein oxidizing the linkage produces an oxophosphoramidate linkage.45. The method of claim 42 , wherein the polynucleotide comprises a sequence of nucleoside subunits complementary to the RNA component of human telomerase claim 42 , and wherein at least two of the nucleoside subunits are joined by a N3′→P5′ phosphoramidate inter-subunit linkage.47. The method of claim 42 , wherein the polynucleotide comprises the sequence TAGGGTTAGACAA.48. The method of claim 47 , wherein all of the internucleotide inter-subunit linkages of the TAGGGTTAGACAA sequence are N3′→P5′ phosphoramidate inter-subunit linkages.52. The method of claim 47 , wherein the C11 nucleotide residue of the TAGGGTTAGACAA sequence derives from a 3′-protected aminonucleoside-5′-phosphoramidite monomer.53. The method of claim 47 , wherein the method ...

Oligonucleotide Compositions and Methods of Making the Same

The present disclosure provides a solid phase method of making oligonucleotides via sequential coupling cycles including at least one coupling of a dinucleotide dimer subunit to a free 3′-terminal group of a growing chain. The oligonucleotides include at least two nucleoside subunits joined by a N3′→P5′ phosphoramidate linkage. The method may include the steps of (a) deprotecting the protected 3′ amino group of a terminal nucleoside attached to a solid phase support, said deprotecting forming a free 3′ amino group; (b) contacting the free 3′ amino group with a 3′-protected amino-dinucleotide-5′-phosphoramidite dimer in the presence of a nucleophilic catalyst to form an internucleoside N3′→P5′ phosphoramidite linkage; and (c) oxidizing (e.g., sulfurizing) the linkage. The compositions produced by the subject methods may include a reduced amount of one or more (N−x) oligonucleotide products. Also provided are pharmaceutical compositions including the subject oligonucleotide compositions.

MORPHOLINO OLIGONUCLEOTIDE MANUFACTURING METHOD

Using a morpholino nucleotide wherein 5′-hydroxy group or a hydroxy group present on the substituent of the 5′-hydroxy group is protected by a protecting group having an alkyl group having not less than 10 and not more than 300 carbon atoms and/or an alkenyl group having not less than 10 and not more than 300 carbon atoms as a starting material, a method capable of efficiently producing the morpholino oligonucleotide in a high yield by a liquid phase synthesis can be provided. 18-. (canceled)9. A method of producing an n+p-mer morpholino oligonucleotide , comprising:(1) condensing (i) a p-mer morpholino oligonucleotide, wherein p is any integer of one or more, and wherein a 5′-hydroxy group is activated (thio)phosphated or activated (thio)phosphoramidated, and a morpholine ring nitrogen atom is protected by a temporary protecting group removable under acidic conditions, with (ii) an n-mer morpholino oligonucleotide, wherein n is an integer of one or more, and wherein a 5′-hydroxy group or, when the 5′-hydroxy group has a substituent having a hydroxy group, the hydroxy group present on the substituent is protected by a protecting group having an alkyl group containing not less than 10 and not more than 300 carbon atoms and/or an alkenyl group containing not less than 10 and not more than 300 carbon atoms, and the morpholine ring nitrogen atom is not protected, by a (thio)phosphoramidate bond or (thio)phosphorodiamidate bond via the morpholine ring nitrogen atom,to obtain a reaction mixture comprising said n+p-mer morpholino oligonucleotide.10. The method according to claim 9 , wherein p is 1.11. The method according to claim 9 , comprising treating said reaction mixture with a quenching agent after completion of said condensing.12. The method according to claim 9 , further comprising:(1′) removing said temporary protecting group of said morpholine ring nitrogen atom by reacting, before said condensing (1), the n-mer morpholino oligonucleotide wherein the 5′-hydroxy ...

A Process for the Preparation of Nucleic Acid by Means of 3'-O-Azidomethyl Nucleotide Triphosphate

The invention relates to a method of nucleic acid synthesis comprising the use of 3′-O-azidomethyl blocked nucleotide triphosphates which comprises the step of adding a capping group to any uncleaved 3′-O-azidomethyl groups and to the use of kits comprising said capping groups in a method of nucleic acid synthesis. The invention also relates to capped nucleotide triphosphates and 3′-O-azidomethyl capping groups.

CYCLIC DINUCLEOTIDES FOR TREATING CONDITIONS ASSOCIATED WITH STING ACTIVITY SUCH AS CANCER

This disclosure features dinucleotide compounds that modulate Stimulator of Interferon Genes (STING) activity, for use for example in the treatment of cancer. This disclosure also features compositions as well as other methods of using and making the same (Formula (A)). A and B are each independently selected from the group consisting of Formulae (i), (ii), (iii), and (iv). 1276-. (canceled)292. A compound selected from{'sup': '6,10', '(1S,3R,6S,8R,9R,10S,12R,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3S,6S,8R,9R,10S,12R,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3S,6S,8R,9R,10S,12S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3R,6S,8R,9R,10S,12S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione; or'}{'sup': '6,10', '(1S,6S,8R,9R,10S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-3,9,12,18-tetrahydroxy-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione.'}293. A pharmaceutical composition comprising a compound according to or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers claim 277 , diluents or excipients.294. A combination pharmaceutical product comprising a compound according to or a pharmaceutically ...

GALNAC PHOSPHORAMIDITES, NUCLEIC ACID CONJUGATES THEREOF AND THEIR USE

This invention generally relates to the field of phosphoramidite derivatives. In particular, the invention relates to N-Acetylgalactosamine phosphoramidite molecules and to conjugates of nucleic acid molecules with N-Acetylgalactosamine containing molecules. Also provided are methods for preparation of these molecules and possible uses thereof, in particular in medicine. 117.-. (canceled)19. Use of a compound according to as a medicament. This application is divisional under 35 U.S.C. § 120 of application Ser. No. 15/517,685 filed Apr. 7, 2017 which is a national stage application under 35 U.S.C. § 371 of PCT Application No. PCT/EP2015/073331 filed Oct. 9, 2015 which claims priority to European Patent Application No. EP14188444.5 filed Oct. 10, 2014 and, European Patent Application No. 15181807.7 filed Aug. 20, 2015, of which each of these applications are hereby incorporated by reference in their entirety.This invention generally relates to the field of phosphoramidite derivatives. In particular, the invention relates to N-Acetylgalactosamine phosphoramidite molecules and to conjugates of nucleic acid molecules with N-Acetylgalactosamine containing molecules. Also provided are methods for preparation of these molecules and possible uses thereof, in particular in medicine.In recent years, approaches have been developed to use nucleic acid molecules in therapy. To favorably influence pharmaceutically relevant properties, the nucleic acid molecules have been conjugated to certain ligands such as peptides, lipids, sterols, and carbohydrates. Nucleic acid conjugates have been extensively evaluated for use in siRNAs, where they are considered essential in order to obtain sufficient in vivo potency. For example, by attachment of a conjugate moiety containing terminal galactose or a derivative thereof to the nucleic acid, thereby targeting the nucleic acid molecule to hepatocytes via binding to the asialoglycoprotein receptor (ASGPR), see for example WO2009/073809, WO2011/ ...

COMPOSITIONS COMPRISING ALTERNATING 2'-MODIFIED NUCLEOSIDES FOR USE IN GENE MODULATION

The present invention provides compositions comprising at least one oligomeric compound comprising an alternating motif and further include a region that is complementary to a nucleic acid target. The compositions are useful for targeting selected nucleic acid molecules and modulating the expression of one or more genes. In preferred embodiments the compositions of the present invention hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. The present invention also provides methods for modulating gene expression. 1105.-. (canceled)106. A composition comprising a first oligomeric compound and a second oligomeric compound , wherein:at least a portion of said first oligomeric compound is capable of hybridizing with at least a portion of said second oligomeric compound;at least a portion of said first oligomeric compound is complementary to and capable of hybridizing to a selected nucleic acid target;{'sub': 1', '2', '1', '2', '3, 'at least one of said first and said second oligomeric compounds comprises a region having the formula X—Y—X, wherein Y is a region of from about 5 to about 12 linked nucleosides and each of Xand Xis, independently, a plurality of linked nucleosides having the formula FSFS, where one of F and S is a 2′-F modified nucleoside and the other of F and S is a 2′-O—CHmodified nucleoside; and'}each of the linked nucleosides is, independently, linked by a phosphodiester or a phosphorothioate internucleoside linkage.107. The composition of wherein said first oligomeric compound comprises a 5′-phosphate group.108. The composition of wherein said second oligomeric compound comprises a 5′-phosphate group.109. The composition of wherein each of said first and said second oligomeric compounds comprise a 5′-phosphate group.110. The composition of wherein at least one of said first and said second oligomeric compounds comprises at least one conjugate group.111. The composition of wherein at least one of said first and ...

Reagents and methods for post-synthetic modification of nucleic acids

The present invention relates to compositions and methods (reagents and protocols) for the post-synthetic modification of nucleic acids obtained from solid-phase oligonucleotide synthesis with a label (such as fluorescent dyes). The coupling reagent is the triazine-based salt 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium (DMT-MM) in the presence of a counteranion.

MODIFIED NUCLEOTIDES

The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula=C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H. 1. (canceled)2. (canceled)3. A modified nucleoside triphosphate molecule comprising a base and a deoxyribose sugar moiety , wherein the 3′ carbon atom of the sugar moiety has covalently attached thereto a group of the structure:{'br': None, '—O—Z'}wherein Z is a removable protecting group comprising an azido group.4. The molecule of claim 3 , wherein the removable protecting group is azidomethyl.5. The molecule of claim 4 , wherein the base is a purine claim 4 , a pyrimidine or a deazapurine.6. The molecule of claim 4 , wherein the base is adenine claim 4 , 7-deazaadenine claim 4 , guanine claim 4 , or 7-deazaguanine.7. The molecule of claim 4 , wherein the base is cytosine claim 4 , thymine or uracil.8. The molecule of claim 4 , wherein the base is linked ...

Dyes and Labeled Molecules

Dimeric and trimeric nucleic acid dyes, and associated systems and methods are provided. Such a dye may form a hairpin-like structure that enables it to stain nucleic acids via a release-on-demand mechanism, for example. Such a dye may have low background fluorescence in the absence of nucleic acids and high fluorescence in the presence of nucleic acids, upon binding therewith, for example. A dye provided herein may be useful in a variety of applications, such as in DNA quantitation in real-time PCR, for example.

METHOD FOR DEBLOCKING OF LABELED OLIGONUCLEOTIDES

The invention relates to a process for deblocking substantially a blocked, detectably labeled oligonucleotide by contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in substantial deblocking of the oligonucleotide, thereby giving the substantially deblocked oligonucleotide. 1. A process for deblocking substantially a blocked , detectably labeled oligonucleotide comprising contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in the deblocking of the oligonucleotide , thereby giving the substantially deblocked oligonucleotide.2. The process of claim 1 , wherein said detectable label is a fluorescent label.3. The process of claim 2 , wherein said detectable label is hexachlorofluorescein.4. The process of claim 2 , wherein said detectable label is DABCYL.5. The process of claim 1 , wherein said nucleophilic amino compound is ammonia.6. The process of claim 5 , wherein said ammonia is present at a psi of about 20 to 200.7. The process of claim 5 , wherein said ammonia is present at a psi of about 80.8. The process of claim 1 , wherein said nucleophilic amino compound is ammonia vapors.9. The process of claim 1 , wherein said nucleophilic amino compound is a Calkylamine.10. The process of claim 1 , further comprising dissolving the substantially deblocked oligonucleotide in a buffer.11. The process of claim 1 , wherein said conditions comprise carrying the process at about room temperature to about 150° C.12. The process of claim 1 , wherein said conditions comprise carrying the process at about 95° C.1314-. (canceled)15. The process of claim 1 , wherein said substantially blocked claim 1 , detectably labeled oligonucleotide is immobilized on a solid phase.16. The process of claim 15 , wherein said substantially deblocked oligonucleotide is released from said solid phase under said ...

Cross-Linked Poly-E-Lysine Non-Particulate Support

The invention provides a non-particulate cross-linked poly-ε-lysine polymer. The poly-ε-lysine and cross linker are linked by amide bonds and may the cross linker has at least two functional groups capable of reacting with an alpha carbon amine of poly-ε-lysine. The polymer is suitably insoluble in water and other solvents and is provided in macro form for example a sheet, article or fibre. The macro form polymer is useful in a wide range of applications including wound treatment, as a medical diagnostic comprising a particulate support and a functional material bound or retained by the support and solid phase synthesis of peptides, oligonucleotides, oligosaccharides, immobilisation of species, cell culturing and in chromatographic separation.

FORMYLPYRROLE-BASED HETEROCYCLES FOR NUCLEIC ACID ATTACHMENT TO SUPPORTS

A compound has Formula I: 2. The compound of claim 1 , wherein the protected aldehyde is selected from the group consisting of an acetal claim 1 , an aminal claim 1 , a dithioacetal claim 1 , a protected hemiaminal claim 1 , an alkene claim 1 , and a protected hemithioacetal.3. The compound of claim 1 , wherein W claim 1 , X claim 1 , Y claim 1 , and Z comprise a fused ring system selected from the group consisting of a benzene claim 1 , a pyridine claim 1 , a furan claim 1 , a thiophene claim 1 , a pyridazine claim 1 , a pyrazine claim 1 , and a pyrimidine.4. The compound of claim 3 , wherein W claim 3 , X claim 3 , Y claim 3 , and Z comprise a fused benzene ring.5. The compound of claim 1 , wherein A is hydrogen or methyl claim 1 , Q is a protected aldehyde claim 1 , Ris N-iPr claim 1 , and Ris OCHCHCN.6. The compound of claim 1 , wherein A is a hydroxyl claim 1 , alkoxy or hydroxyalkyl claim 1 , Q is a protected aldehyde claim 1 , Ris N-iPr claim 1 , and Ris OCHCHCN.8. The compound of claim 7 , wherein the protected aldehyde is selected from the group consisting of an acetal claim 7 , an aminal claim 7 , a dithioacetal claim 7 , a protected hemiaminal claim 7 , an alkene claim 7 , and a protected hemithioacetal.9. The compound of wherein W claim 7 , X claim 7 , Y claim 7 , and Z comprise a fused ring system selected from the group consisting of a benzene claim 7 , a pyridine claim 7 , a furan claim 7 , a thiophene claim 7 , a pyridazine claim 7 , a pyrazine claim 7 , and a pyrimidine.10. The compound of claim 9 , wherein W claim 9 , X claim 9 , Y claim 9 , and Z comprise a fused benzene ring.11. The compound of claim 7 , wherein A is hydrogen or methyl claim 7 , Q is a protected aldehyde claim 7 , Nu is selected from the group consisting of a 3′-phosphate-linked nucleic acid claim 7 , a 3′-thiophosphate-linked nucleic acid claim 7 , and a 3′-phosphate linked modified nucleic acid.12. The compound of claim 7 , wherein A is hydrogen or methyl claim 7 , Q is a ...

Modified nucleotides

The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

Enrichment and Sequencing of RNA Species

Provided herein is a method for making an cDNA library, comprising adding an affinity tag-labeled GMP to the 5′ end of targeted RNA species in a sample by optionally decapping followed by incubating the sample with an affinity tag-labeled GTP and a capping enzyme, enriching for RNA comprising the affinity tag-labeled GMP using an affinity matrix that binds to the affinity tag, reverse transcribing the enriched RNA to produce a population of cDNAs, and adding a tail to the 3′ end of the population of cDNAs using a terminal transferase, to produce an cDNA library.

CYCLIC DINUCLEOTIDES FOR TREATING CONDITIONS ASSOCIATED WITH STING ACTIVITY SUCH AS CANCER

This disclosure features dinucleotide compounds that modulate Stimulator of Interferon Genes (STING) activity, for use for example in the treatment of cancer. This disclosure also features compositions as well as other methods of using and making the same. A and B are each independently selected from the group consisting of Formulae (i), (ii), (iii), and (iv): 1145-. (canceled)147. The compound according to wherein X and X′ are each O.154. The compound according to wherein each of X claim 146 , X claim 146 , Xand Xis O.157. A pharmaceutical composition comprising a compound according to or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers claim 146 , diluents or excipients.158. A combination pharmaceutical product comprising a compound according to or a pharmaceutically acceptable salt thereof together with one or more other therapeutically active agents.159. A method for modulating STING activity comprising contacting STING with a compound according to .160. A method of treating cancer claim 146 , comprising administering to a subject in need of such treatment an effective amount of a compound according to .161. The method of wherein the compound is administered in combination with one or more additional cancer therapies.162. The method of wherein the one or more additional cancer therapies comprises surgery claim 161 , radiotherapy claim 161 , chemotherapy claim 161 , toxin therapy claim 161 , immunotherapy claim 161 , cryotherapy or gene therapy claim 161 , or a combination thereof.163. A method for treating cancer in a subject in need thereof claim 146 , comprising administering an effective amount of a compound claim 146 , according to claim 146 , or a pharmaceutically acceptable salt thereof claim 146 ,in combination with the administration of a therapeutically effective amount of one or more immuno-oncology agents.164. The method of claim 163 , wherein the immuno-oncology agent is a anti-PD-1 antibody.165. The ...

MISPRIMING PREVENTION REAGENTS

Provided herein are mispriming prevention reagents, compositions and kits comprising such reagents and methods of use thereof. 1162-. (canceled)163. A method of creating a cDNA comprising: (i) RNA;', '(iv) a reverse transcriptase;', '(v) dNTPs; and', (1) a first condition-dependent stem region comprising a 5′ terminal covalently linked moiety and a first stem nucleic acid sequence, wherein the first stem nucleic acid sequence is at least 6 nucleotides in length and wherein the 5′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion;', '(2) a condition-dependent loop region comprising a loop nucleic acid sequence of at least 3 nucleotides in length; and', '(3) a second condition-dependent stem region comprising a second stem nucleic acid sequence and a 3′ terminal covalently linked moiety, wherein the second stem nucleic acid sequence is at least 6 nucleotides in length and is complementary to the first stem nucleic acid sequence, wherein the 3′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion, wherein the 3′ terminal covalently linked moiety is non-identical to the 5′ terminal covalently linked moiety, wherein the 3′ terminus of the second condition-dependent stem region is non-extendable by the reverse transcriptase or by a thermostable DNA polymerase, wherein the second condition-dependent stem region hybridizes to the first condition-dependent stem region with a stem melting temperature that is no greater than the first primer melting temperature and the second primer melting temperature, and wherein hybridization of the second condition-dependent stem region to the first condition-dependent stem region causes the reagent to acquire a stem-loop hairpin conformation; and, '(vi) a mispriming prevention reagent comprising a nucleic acid molecule comprising, in 5′ to 3′ order], '(a) forming a reaction mixture comprising(b) incubating the ...

SENSITIVE OLIGONUCLEOTIDE SYNTHESIS USING SULFUR-BASED FUNCTIONS AS PROTECTING GROUPS AND LINKERS

Embodiments for the synthesis of sensitive oligonucleotides as well as insensitive oligonucleotides are provided. Sulfur-based groups are used for the protection of exo-amino groups of nucleobases, phosphate groups and 2′-OH groups, and as cleavable linker for linking oligonucleotides to a support. Oligonucleotide syntheses are achieved under typical conditions using phosphoramidite chemistry with important modifications. To prevent replacing sulfur-based protecting groups by acyl groups via cap-exchange, special capping agents are used. To retain hydrophobic tag to assist RP HPLC purification, special phosphoramidites are used in the last synthetic cycle. With the sulfur-based groups for protection and linking, oligonucleotide deprotection and cleavage are achieved via oxidation followed by beta-elimination under mild conditions. Therefore, besides for insensitive oligonucleotide synthesis, the embodiments of the invention are capable for the synthesis of oligonucleotide analogs containing sensitive functional groups that cannot survive the harsh conditions used in prior art oligonucleotide synthesis technologies. 2. Derivatized nucleoside phosphoramidites of wherein Rand Rare —SRwith Rbeing independent alkyl claim 1 , derivatized alkyl claim 1 , aryl and derivatized aryl; or R-R=—S[C(R)R]S— claim 1 , —S{[C(R)R]o[C(R)R]}S— or —S{[C(R)R]S[C(R)R]}S— wherein independently Rand Rare H atoms or alkyl groups independently in the repetitions claim 1 , and m claim 1 , n and p are independent integers.3. Derivatized nucleoside phosphoramidites of wherein Ris selected from (II-IV); R-R=—S[C(R)R]S— claim 1 , —S{[C(R)R]O[C(R)R]}S— or —S{[C(R)R]S[C(R)R]}S— wherein independently Rand Rare H or alkyl groups independently in the repetitions claim 1 , and m claim 1 , n and p are independent integers; and Ris an alkyl group with less than six carbons claim 1 , and R=H.4. Derivatized nucleoside phosphoramidites of wherein Ris selected from (II-IV); R-R=—S[C(R)R]S— claim 1 , —S{[C(R)R ...

Oligonucleotide Analogues Incorporating 5-Aza-Cytosine Therein

Oligonucleotide analogues are provided that incorporate 5-aza-cytosine in the oligonucleotide sequence, e.g., in the form of 5-aza-2′-deoxycytidine (decitabine) or 5-aza-cytidine. In particular, oligonucleotide analogues rich in decitabine-deoxyguanosine islets (DpG and GpD) are provided to target the CpG islets in the human genome, especially in the promoter regions of genes susceptible to aberrant hypermethylation. Such analogues can be used for modulation of DNA methylation, such as effective inhibition of methylation of cytosine at the C-5 position. Methods for synthesizing these oligonucleotide analogues and for modulating nucleic acid methylation are provided. Also provided are phosphoramidite building blocks for synthesizing the oligonucleotide analogues, methods for synthesizing, formulating and administering these compounds or compositions to treat conditions, such as cancer and hematological disorders. 1114-. (canceled)115. A method of treating a condition , the method comprising administering to a subject in need thereof a therapeutically-effective amount of a dinucleotide analogue , or a pharmaceutically-acceptable salt thereof , wherein the dinucleotide analogue , or the pharmaceutically-acceptable salt thereof , comprises a phospholinker , wherein the number of phosphorus atoms in the phospholinker is one , wherein the linker is not a phosphorothioate linker.116. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a 5-aza-cytosine group.117. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a decitabine group.118. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a deoxyguanosine group.119. The method of claim 115 , wherein the dinucleotide analogue is the pharmaceutically-acceptable salt claim 115 , wherein the pharmaceutically-acceptable salt is a ...

COMPOUNDS,COMPOSITIONS, AND METHODS FOR THE TREATMENT OF DISEASE

Disclosed are compounds and compositions for the activation or induction of expression of a pattern recognition receptor (e.g., STING, RIG-I, MDA5), and methods of use thereof. 146-. (canceled)54. A method of treating cancer claim 47 , comprising administering to a subject in need thereof an effective amount of a compound of or a pharmaceutically acceptable salt thereof.61. The method of claim 54 , wherein the cancer is a cancer of the breast claim 54 , bone claim 54 , brain claim 54 , cervix claim 54 , colon claim 54 , gastrointestinal tract claim 54 , eye claim 54 , gall bladder claim 54 , lymph nodes claim 54 , blood claim 54 , lung claim 54 , liver claim 54 , skin claim 54 , mouth claim 54 , prostate claim 54 , ovary claim 54 , penis claim 54 , pancreas claim 54 , uterus claim 54 , testicles claim 54 , stomach claim 54 , thymus claim 54 , or thyroid.62. The method of claim 54 , wherein the cancer is acute monocytic leukemia claim 54 , lymphoma claim 54 , melanoma claim 54 , colon cancer claim 54 , or breast cancer. This application claims the benefit of priority to U.S. provisional patent application Nos. 62/359,039, filed Jul. 6, 2016; 62/363,118, filed Jul. 15, 2016; 62/403,530, filed Oct. 3, 2016; 62/411,424, filed Oct. 21, 2016; 62/444,141, filed Jan. 9, 2017; 62/462,679, filed Feb. 23, 2017; 62/470,746, filed Mar. 13, 2017; and 62/508,846, May 19, 2017; the contents of each of which are hereby incorporated by reference in their entireties.This disclosure relates to compounds and compositions that activate the innate immune defense system and induce expression of pattern recognition receptors in a host, as well as methods of use for the treatment of a proliferative disease (e.g., cancer).A key feature of the innate immune system is the recognition and elimination of foreign substances. Identification of these pathogenic invaders occurs through host recognition of evolutionarily conserved microbial structures known as pathogen-associated molecular patterns ( ...

Compositions and method for synthesizing nucleic acids

Provided herein are compositions, kits and methods for synthesis of nucleic acids. Also provided herein are compositions and methods for synthesizing strands of nucleic acid across different nucleic acid back-bones hybridized together using a strand displacing polymerase.

BIOPARTICLE ISOLATION AND THERAPEUTIC APPLICATION THEREOF

Compositions and methods for the isolation of protein-nucleic acid complexes, extracellular vesicle (EV) (e.g., microvesicles) and free nucleic acids (collectively referred to as “bioparticles”) released by mammalian cells into body fluids or cell culture media are provided. Isolated bioparticles of the current disclosure contain biomolecules that are useful as diagnostic/prognostic biomarkers or for identification of therapeutic targets (e.g., disease or disorder-associated miRNAs, circulating tumor DNA). Isolation of biomolecules results in purification and concentration. Methods for producing biofluids without detectable bioparticles, largely depleted of bioparticles, and/or possessing a reduced concentration of bioparticles compared to a biofluid starting material (collectively termed “bioparticle-depleted”) are provided. Bioparticle-depleted biofluid is useful, e.g., in experimental systems where desirable to obtain a biofluid lacking or substantially depleted of endogenous bioparticles from the source material. Non-toxic bioparticle absorbing materials (e.g., exosome-reducing materials) can also be used for prophylactic, therapeutic, validation and/or experimental purposes. 1. A method for isolating , amplifying or both isolating and amplifying cell-free nucleic acids from a liquid sample with enhanced efficiency comprising:a) obtaining a liquid sample from a subject or cell culture;b) contacting said liquid sample with a crystallizing agent under conditions suitable to allow for crystal formation, thereby creating an admixture;c) incubating said admixture for a period of time sufficient to allow for crystal formation;d) separating said admixture to obtain a particle fraction containing bioparticles; ande) isolating amplifying or both isolating and amplifying cell-free nucleic acids from the particle fraction containing bioparticles, thereby isolating, amplifying or both isolating and amplifying cell-free nucleic acids from the liquid sample with enhanced ...