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RU2018131783A - Транс-реплицирующая рнк - Google Patents

Транс-реплицирующая рнк Download PDF

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RU2018131783A
RU2018131783A RU2018131783A RU2018131783A RU2018131783A RU 2018131783 A RU2018131783 A RU 2018131783A RU 2018131783 A RU2018131783 A RU 2018131783A RU 2018131783 A RU2018131783 A RU 2018131783A RU 2018131783 A RU2018131783 A RU 2018131783A
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alphavirus
replicase
rna
expression
replication
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RU2018131783A
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RU2752580C2 (ru
RU2018131783A3 (ru
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Тим БАЙССЕРТ
Угур ЗАХИН
Марио ПЕРКОВИЧ
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Бионтэк Рна Фармасьютикалз Гмбх
Трон - Транслационале Онкологи Ан Дер Униферзитетсмедицин Дер Иоганн Гутенберг-Униферзитет Майнц Гемайннютциге Гмбх
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Claims (42)

1. Система, содержащая:
конструкцию РНК для экспрессии альфавирусной репликазы,
РНК-репликон, который может быть реплицирован репликазой in trans,
причем конструкция РНК для экспрессии альфавирусной репликазы содержит 5’-кэп для трансляции репликазы.
2. Система по п. 1, в которой 5’-кэп представляет собой природный 5’-кэп или аналог 5’-кэпа.
3. Система по п. 1 или 2, в которой конструкция РНК для экспрессии альфавирусной репликазы не содержит элемент участка внутренней посадки рибосомы (IRES) для трансляции репликазы.
4. Система по любому из пп. 1-3, в которой конструкция РНК для экспрессии альфавирусной репликазы содержит:
(1) 5' UTR,
(2) открытую рамку считывания, кодирующую репликазу, и
(3) 3' UTR.
5. Система по п. 4, в которой 5' UTR и/или 3' UTR являются ненативными для альфавируса, из которого получена репликаза.
6. Система по любому из пп. 1-5, в которой открытая рамка считывания, кодирующая альфавирусную репликазу, содержит кодирующую область(-и) для неструктурных белков, необходимых для репликации РНК.
7. Система по любому из пп. 1-6, в которой конструкция РНК для экспрессии альфавирусной репликазы содержит 3' поли(А)-последовательность.
8. Система по любому из пп. 1-7, в которой конструкция РНК для экспрессии альфавирусной репликазы не может быть реплицирована репликазой.
9. Система по любому из пп. 1-8, в которой РНК-репликон содержит:
(1) 5' репликационную последовательность распознавания альфавируса и
(2) 3' репликационную последовательность распознавания альфавируса.
10. Система по п. 9, в которой 5' репликационная последовательность распознавания альфавируса и 3' репликационная последовательность распознавания альфавируса направляют репликацию РНК-репликона в присутствии репликазы.
11. Система по п. 9 или 10, в которой 5' репликационная последовательность распознавания альфавируса и 3' репликационная последовательность распознавания альфавируса являются нативными для альфавируса, из которого получена репликаза.
12. Система по любому из пп. 1-11, в которой РНК-репликон содержит гетерологичную нуклеиновую кислоту.
13. Система по любому из пп. 1-12, в которой РНК-репликон содержит открытую рамку считывания, кодирующую представляющий интерес белок.
14. Система по п. 13, в которой открытая рамка считывания, кодирующая представляющий интерес белок, является ненативной для альфавируса, из которого получена репликаза.
15. Система по п. 13 или 14, в которой экспрессия открытой рамки считывания, кодирующей представляющий интерес белок, находится под контролем субгеномного промотора.
16. Система по п. 15, в которой субгеномный промотор является нативным для альфавируса, из которого получена репликаза.
17. Система по п. 15 или 16, в которой субгеномный промотор представляет собой промотор структурного белка альфавируса.
18. Система по любому из пп. 1-17, в которой РНК-репликон содержит 3' поли(А)-последовательность.
19. Система по любому из пп. 1-18, в которой РНК-репликон содержит 5’-кэп.
20. Система по любому из пп. 1-19, в которой конструкция РНК для экспрессии альфавирусной репликазы и/или РНК-репликон не содержит открытую рамку считывания, кодирующую интактный структурный белок альфавируса.
21.     Система по любому из пп. 1-20, в которой альфавирус представляет собой вирус леса Семлики или вирус венесуэльского лошадиного энцефалита, или вирус Синдбис, или вирус Чикунгунья.
22. Конструкция РНК для экспрессии альфавирусной репликазы, содержащая 5’-кэп для трансляции репликазы.
23. ДНК, содержащая последовательность нуклеиновой кислоты, кодирующую конструкцию РНК для экспрессии альфавирусной репликазы, РНК-репликон или обе системы по любому из пп. 1-21.
24. Способ получения белка в клетке, предусматривающий следующие стадии:
(а) получение конструкции РНК для экспрессии альфавирусной репликазы,
(b) получение РНК-репликона, который может быть реплицирован репликазой in trans и содержит открытую рамку считывания, кодирующую белок, и
(c) инокуляция клетки смесью конструкции РНК для экспрессии альфавирусной репликазы и РНК-репликона,
причем конструкция РНК для экспрессии альфавирусной репликазы содержит 5’-кэп для трансляции репликазы.
25. Клетка, инокулированная по способу по п. 24.
26. Способ получения белка у субъекта, предусматривающий следующие стадии:
(а) получение конструкции РНК для экспрессии альфавирусной репликазы,
(b) получение РНК-репликона, который может быть реплицирован репликазой in trans и содержит открытую рамку считывания, кодирующую белок, и
(c) введение конструкции РНК для экспрессии альфавирусной репликазы и РНК-репликона субъекту,
причем конструкция РНК для экспрессии альфавирусной репликазы содержит 5’-кэп для трансляции репликазы.
RU2018131783A 2016-03-21 2017-03-13 Транс-реплицирующая рнк RU2752580C2 (ru)

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PCT/EP2016/056160 WO2017162265A1 (en) 2016-03-21 2016-03-21 Trans-replicating rna
EPPCT/EP2016/056160 2016-03-21
PCT/EP2017/055813 WO2017162461A1 (en) 2016-03-21 2017-03-13 Trans-replicating rna

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RU2752580C2 RU2752580C2 (ru) 2021-07-29

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