EA201991441A1 - ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9 - Google Patents

ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9

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Publication number
EA201991441A1
EA201991441A1 EA201991441A EA201991441A EA201991441A1 EA 201991441 A1 EA201991441 A1 EA 201991441A1 EA 201991441 A EA201991441 A EA 201991441A EA 201991441 A EA201991441 A EA 201991441A EA 201991441 A1 EA201991441 A1 EA 201991441A1
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Prior art keywords
thermocas9
cell
gene
polynucleotide encoding
interest
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EA201991441A
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English (en)
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Джон Ван Дер Ост
Рихард Ван Краненбург
Эллеке Фенна Босма
Иоаннис Маугиакос
Прартхана Моханраджу
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Вагенинген Университейт
Стихтинг Вор Де Технише Ветенсхаппен
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Priority claimed from PCT/EP2017/082870 external-priority patent/WO2018109101A1/en
Publication of EA201991441A1 publication Critical patent/EA201991441A1/ru

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Abstract

Полинуклеотид, кодирующий белок ThermoCas9 из Geobacillus thermodenitrificans, и конститутивный промотор используют для генетической модификации эукариотических клеток, например грибов, дрожжей или водорослей, таким образом, что эндонуклеаза ThermoCas9 интегрируется и экспрессируется с генома клетки. Затем используют вторую экспрессионную плазмиду для трансфицирования этих экспрессирующих ThermoCas9 клеток, при этом вторая плазмида содержит индуцируемый промотор и полинуклеотид, кодирующий гид-РНК. Гид-РНК в сочетании с ThermoCas9 обеспечивают направленную эндонуклеазную активность для расщепления ДНК клетки в желаемом локусе или интересующем гене. В клетку также вводят репарационный олигонуклеотид, за счет чего после расщепления ДНК в клетке происходит гомологичная рекомбинация с репарационным олигонуклеотидом, вследствие чего имеет место либо делеция, либо замена нуклеотидов в локусе или интересующем гене. Описаны экспрессионные векторы и способы использования векторов для осуществления опосредованного ThermoCas9 редактирования генов при повышенных температурах, например превышающих 30°C.
EA201991441A 2016-12-14 2017-12-14 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9 EA201991441A1 (ru)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
PCT/EP2016/081077 WO2018108272A1 (en) 2016-12-14 2016-12-14 Thermostable cas9 nucleases
PCT/EP2017/070796 WO2018108338A1 (en) 2016-12-14 2017-08-16 Thermostable cas9 nucleases
PCT/EP2017/082870 WO2018109101A1 (en) 2016-12-14 2017-12-14 Thermostable cas9 nucleases

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EA201991441A1 true EA201991441A1 (ru) 2020-01-09

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EA201991443A EA201991443A1 (ru) 2016-12-14 2016-12-14 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9
EA201991442A EA201991442A1 (ru) 2016-12-14 2017-08-16 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9
EA201991440A EA201991440A1 (ru) 2016-12-14 2017-08-16 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9
EA201991441A EA201991441A1 (ru) 2016-12-14 2017-12-14 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9

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EA201991443A EA201991443A1 (ru) 2016-12-14 2016-12-14 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9
EA201991442A EA201991442A1 (ru) 2016-12-14 2017-08-16 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9
EA201991440A EA201991440A1 (ru) 2016-12-14 2017-08-16 ТЕРМОСТАБИЛЬНЫЕ НУКЛЕАЗЫ Cas9

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US (5) US11242513B2 (ru)
EP (4) EP3555275A1 (ru)
JP (6) JP7182545B2 (ru)
KR (4) KR20190104342A (ru)
CN (4) CN110312792B (ru)
AU (4) AU2016432443B2 (ru)
BR (4) BR112019012155A2 (ru)
CA (4) CA3046824A1 (ru)
EA (4) EA201991443A1 (ru)
PH (4) PH12019501335A1 (ru)
WO (3) WO2018108272A1 (ru)

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