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RU2018136614A - Отличные от человека животные с гуманизированным геном сигнального регуляторного белка - Google Patents

Отличные от человека животные с гуманизированным геном сигнального регуляторного белка Download PDF

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RU2018136614A
RU2018136614A RU2018136614A RU2018136614A RU2018136614A RU 2018136614 A RU2018136614 A RU 2018136614A RU 2018136614 A RU2018136614 A RU 2018136614A RU 2018136614 A RU2018136614 A RU 2018136614A RU 2018136614 A RU2018136614 A RU 2018136614A
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rodent
sirpα
gene
humanized
human
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RU2018136614A
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English (en)
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Эндрю Дж. МЕРФИ
О. Гэвин Терстон
Бинду ВАРГХЕСЕ
Каган ГУРЕР
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Регенерон Фармасьютикалс, Инк.
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Publication of RU2018136614A publication Critical patent/RU2018136614A/ru

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Claims (39)

1. Грызун для экспрессии гуманизированного белка SIRPα,
причем геном грызуна содержит замещение экзонов 2, 3 и 4 гена SIRPα грызуна в эндогенном локусе SIRPα грызуна на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα,
причем указанный гуманизированный ген SIRPα функционально связан с промотором SIRPα грызуна в указанном эндогенном локусе SIRPα грызуна и экспрессирует у указанного грызуна гуманизированный белок SIRPα, содержащий внеклеточную часть белка SIRPα человека, кодируемого указанным геном SIRPα человека, и внутриклеточную часть белка SIRPα грызуна, кодируемого указанным геном SIRPα грызуна, и
причем грызуном является крыса.
2. Грызун по п. 1, у которого гуманизированный ген SIRPα содержит экзоны 1, 5, 6, 7 и 8 указанного гена SIRPα грызуна.
3. Грызун по п. 1 или 2, причем указанный грызун не экспрессирует белок SIRPα грызуна.
4. Выделенная клетка грызуна для экспрессии гуманизированного белка SIRPα,
причем геном выделенной клетки грызуна содержит замещение экзонов 2, 3 и 4 гена SIRPα грызуна в эндогенном локусе SIRPα грызуна на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα,
причем указанный гуманизированный ген SIRPα функционально связан с промотором SIRPα грызуна в указанном эндогенном локусе SIRPα грызуна и кодирует гуманизированный белок SIRPα, содержащий внеклеточную часть белка SIRPα человека, кодируемого указанным геном SIRPα человека, и внутриклеточную часть белка SIRPα грызуна, кодируемого указанным геном SIRPα грызуна, и
причем выделенной клеткой грызуна является клетка крысы.
5. Выделенная клетка грызуна по п. 4, у которой гуманизированный ген SIRPα содержит экзоны 1, 5, 6, 7 и 8 указанного гена SIRPα грызуна.
6. Выделенная клетка грызуна по п. 4 или п. 5, причем указанная клетка или ткань не экспрессирует белок SIRPα грызуна.
7. Выделенная ткань грызуна для экспрессии гуманизированного белка SIRPα,
причем геном выделенной ткани грызуна содержит замещение экзонов 2, 3 и 4 гена SIRPα грызуна в эндогенном локусе SIRPα грызуна на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα,
причем указанный гуманизированный ген SIRPα функционально связан с промотором SIRPα грызуна в указанном эндогенном локусе SIRPα грызуна и кодирует гуманизированный белок SIRPα, содержащий внеклеточную часть белка SIRPα человека, кодируемого указанным геном SIRPα человека, и внутриклеточную часть белка SIRPα грызуна, кодируемого указанным геном SIRPα грызуна, и
причем выделенной тканью грызуна является выделенная ткань крысы.
8. Выделенная ткань грызуна по п. 7, у которой гуманизированный ген SIRPα содержит экзоны 1, 5, 6, 7 и 8 указанного гена SIRPα грызуна.
9. Выделенная ткань грызуна по п. 7 или 8, причем клетка или ткань не экспрессирует белок SIRPα грызуна.
10. Способ получения грызуна для экспрессии гуманизированного белка SIRPα, предусматривающий
(а) замещение экзонов 2, 3 и 4 гена SIRPα грызуна в эндогенном локусе SIRPα грызуна у эмбриональной стволовой (ЭС) клетки грызуна на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα, причем указанный гуманизированный ген SIRPα функционально связан с промотором SIRPα грызуна в указанном эндогенном локусе SIRPα грызуна и кодирует гуманизированный белок SIRPα, содержащий внеклеточную часть белка SIRPα человека, кодируемого указанным геном SIRPα человека, и внутриклеточную часть белка SIRPα грызуна, кодируемого указанным геном SIRPα грызуна, таким образом, получая модифицированную ЭС клетку грызуна, содержащую гуманизированный ген SIRPα, причем ЭС клеткой грызуна является ЭС клетка крысы; и
(b) получение грызуна с использованием модифицированной ЭС клетки, полученной на стадии (а), причем грызуном является крыса.
11. Способ трансплантации грызуну человеческих гематопоэтических клеток, предусматривающий стадии:
(a) получения грызуна по любому из пп. 1-3 и
(b) трансплантации грызуну одной или нескольких человеческих гематопоэтических клеток.
12. Способ фагоцитоза меченого субстрата клетками, предусматривающий
(а) получение одной или нескольких клеток грызуна, геном которых содержит замещение экзонов 2, 3 и 4 гена SIRPα грызуна в эндогенном локусе SIRPα грызуна на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα,
причем указанный гуманизированный ген SIRPα функционально связан с промотором SIRPα грызуна в указанном эндогенном локусе SIRPα грызуна и кодирует гуманизированный белок SIRPα, содержащий внеклеточную часть белка SIRPα человека, кодируемого указанным геном SIRPα человека, и внутриклеточную часть белка SIRPα грызуна, кодируемого указанным геном SIRPα грызуна, и
причем одной или несколькими клетками грызуна являются клетки крысы;
(b) инкубацию одной или нескольких клеток, полученных на стадии (а), с меченым субстратом и
(c) измерение фагоцитоза меченого субстрата одной или несколькими клетками, упомянутыми на стадии (b).
13. Способ измерения фагоцитоза антигена клетками, предусматривающий
(a) получение грызуна по любому из пп. 1-3;
(b) воздействие на грызуна антигеном и
(c) измерение фагоцитоза антигена одной или несколькими клетками грызуна.
14. Способ оценки терапевтической эффективности потенциального лекарственного средства, направленно воздействующего на человеческие злокачественные клетки, предусматривающий
(a) получение грызуна по любому из пп. 1-3;
(b) трансплантацию грызуну одной или нескольких человеческих злокачественных клеток;
(c) введение указанному грызуну потенциального лекарственного средства, направленно воздействующего на человеческие злокачественные клетки; и
(d) мониторинг человеческих злокачественных клеток у грызуна для определения терапевтической эффективности потенциального лекарственного средства.
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