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RU2018122288A - Композиции и способы с участием нуклеиновых кислот, нацеленных на нуклеиновые кислоты - Google Patents

Композиции и способы с участием нуклеиновых кислот, нацеленных на нуклеиновые кислоты Download PDF

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Publication number
RU2018122288A
RU2018122288A RU2018122288A RU2018122288A RU2018122288A RU 2018122288 A RU2018122288 A RU 2018122288A RU 2018122288 A RU2018122288 A RU 2018122288A RU 2018122288 A RU2018122288 A RU 2018122288A RU 2018122288 A RU2018122288 A RU 2018122288A
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Prior art keywords
cas9 protein
crispr cas9
modified crispr
protein according
cell
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RU2018122288A
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RU2018122288A3 (ru
Inventor
Эндрю Пол МЭЙ
Рейчел И. ОРВИТЦ
Дженнифер А. ДУДНА
Джеймс М. БЕРГЕР
Мэттью Мэррилл КАРТЕР
Пол ДОНОХЬЮ
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Карибо Биосайенсиз, Инк.
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Publication of RU2018122288A publication Critical patent/RU2018122288A/ru
Publication of RU2018122288A3 publication Critical patent/RU2018122288A3/ru

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Claims (21)

1. Модифицированный белок CRISPR Cas9, который содержит модификацию в участке белка CRISPR Cas9, соответствующем положениям остатков аминокислот 170-312 белка Cas9 из S. pyogenes- (SEQ ID NO: 8), причем указанная модификация включает вставку, делецию или их комбинацию.
2. Модифицированный белок CRISPR Cas9 по п. 1, отличающийся тем, что указанная модификация находится в участках обнаруженного сходства последовательностей между указанным белком Cas9 и белком Cas9 из S. pyogenes- (SEQ ID NO: 8).
3. Модифицированный белок CRISPR Cas9 по п. 1 или 2, отличающийся тем, что указанная модификация включает делецию.
4. Модифицированный белок CRISPR Cas9 по п. 3, отличающийся тем, что указанная модификация включает делецию более чем 5 остатков аминокислот.
5. Модифицированный белок CRISPR Cas9 по п. 3, отличающийся тем, что указанная модификация включает делецию более чем 100 остатков.
6. Модифицированный белок CRISPR Cas9 по любому из пп. 1-5, отличающийся тем, что указанная модификация включает делецию, а также вставку последовательности неприродных аминокислот.
7. Модифицированный белок CRISPR Cas9 по п. 1 или 2, отличающийся тем, что указанная модификация включает вставку последовательности неприродных аминокислот с образованием гибридного белка.
8. Модифицированный белок CRISPR Cas9 по п. 7, отличающийся тем, что указанная последовательность неприродных аминокислот содержит функциональный домен.
9. Модифицированный белок CRISPR Cas9 по п. 1 или 2, отличающийся тем, что указанный модифицированный белок CRISPR Cas9 представляет собой гибридный белок.
10. Модифицированный белок CRISPR Cas9 по п. 9, отличающийся тем, что указанный модифицированный гибридный белок CRISPR Cas9 содержит детектируемую метку.
11. Модифицированный белок CRISPR Cas9 по п. 9, отличающийся тем, что указанный модифицированный гибридный белок CRISPR Cas9 содержит модифицированный белок CRISPR Cas9 и нуклеазу.
12. Модифицированный белок CRISPR Cas9 по п. 9, отличающийся тем, что указанная нуклеаза представляет собой FokI.
13. Модифицированный белок CRISPR Cas9 по любому из пп. 1-12, отличающийся тем, что указанный модифицированный белок CRISPR Cas9 обладает ферментативной активностью.
14. Модифицированный белок CRISPR Cas9 по любому из пп. 1-13, отличающийся тем, что указанный модифицированный белок CRISPR Cas9 подвергнут оптимизации кодонов для представляющей интерес клетки-хозяина.
15. Модифицированный белок CRISPR Cas9 по п. 14, отличающийся тем, что указанная клетка-хозяин представляет собой бактериальную клетку, клетку архей, клетку одноклеточного эукариотического организма, растительную клетку, клетку водорослей, клетку гриба, клетку животного, клетку беспозвоночного животного, клетку позвоночного животного, клетку млекопитающего.
16. Модифицированный белок CRISPR Cas9 по п. 15, отличающийся тем, что указанная клетка-хозяин представляет собой клетку человека.
17. Полинуклеотид, кодирующий модифицированный белок CRISPR Cas9 по любому из пп. 1-16.
18. Комплекс, включающий модифицированный белок CRISPR Cas9 по любому из пп. 1-16 и нуклеиновая кислота для нацеливания на нуклеиновую кислоту.
19. Набор, содержащий:
модифицированный белок CRISPR Cas9 по любому из пп. 1-16, или полинуклеотид, который кодирует указанный модифицированный белок CRISPR Cas9; и реагент.
20. Набор по п. 19, дополнительно содержащий нуклеиновую кислоту для нацеливания на нуклеиновую кислоту или полинуклеотид, кодирующий нуклеиновую кислоту для нацеливания на нуклеиновую кислоту.
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