RU2020103731A - Автоматизированные способы обработки клеток, модули, инструменты и системы - Google Patents
Автоматизированные способы обработки клеток, модули, инструменты и системы Download PDFInfo
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- 150000007523 nucleic acids Chemical class 0.000 claims 24
- 102000039446 nucleic acids Human genes 0.000 claims 24
- 108020004707 nucleic acids Proteins 0.000 claims 24
- 230000009466 transformation Effects 0.000 claims 11
- 239000003153 chemical reaction reagent Substances 0.000 claims 10
- 239000012530 fluid Substances 0.000 claims 7
- 238000004520 electroporation Methods 0.000 claims 4
- 238000001914 filtration Methods 0.000 claims 4
- 230000003287 optical effect Effects 0.000 claims 3
- 238000011084 recovery Methods 0.000 claims 3
- 101710163270 Nuclease Proteins 0.000 claims 2
- 230000010307 cell transformation Effects 0.000 claims 1
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- 238000010362 genome editing Methods 0.000 claims 1
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Claims (47)
1. Автоматизированный многомодульный инструмент для редактирования клеток, содержащий:
корпус, выполненный с возможностью размещения всех или некоторых модулей;
емкость, выполненную с возможностью приема клеток;
одну или более емкостей, выполненных с возможностью приема нуклеиновых кислот;
модуль трансформации, выполненный с возможностью введения нуклеиновых кислот в клетки;
модуль восстановления, выполненный с возможностью восстановления клеток после трансформации клеток в модуле трансформации;
модуль направляемого нуклеазой редактирования, выполненный с возможностью редактирования нуклеиновых кислот в клетках с помощью введенных нуклеиновых кислот;
процессор, выполненный с возможностью обеспечения работы автоматизированного многомодульного инструмента для редактирования клеток, основываясь на вводе данных пользователем и/или выборе пользователем предварительно подготовленного скрипта, и
автоматизированную систему подачи жидкости для перемещения жидкостей непосредственно из одного из: модуля для выращивания, модуля трансформации или модуля направляемого нуклеазой редактирования в другой из: модуля для выращивания, модуля трансформации или модуля направляемого нуклеазой редактирования без вмешательства пользователя.
2. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, в котором нуклеиновые кислоты в одной или более емкостях содержат остов и редактирующую кассету, причем автоматизированное многомодульное устройство для редактирования клеток дополнительно содержит модуль сборки нуклеиновых кислот.
3. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, в котором автоматизированная система подачи жидкости содержит сиппер или дозатор.
4. Автоматизированный многомодульный инструмент для редактирования клеток по п. 2, в котором модуль сборки нуклеиновых кислот выполнен с возможностью осуществления изотермической сборки нуклеиновых кислот.
5. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, в котором модуль редактирования и модуль восстановления объединены в один модуль.
6. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, дополнительно содержащий модуль для выращивания, выполненный с возможностью выращивания клеток.
7. Автоматизированный многомодульный инструмент для редактирования клеток по п. 6, в котором модуль для выращивания измеряет оптическую плотность выращиваемых клеток.
8. Автоматизированный многомодульный инструмент для редактирования клеток по п. 7, в котором оптическую плотность измеряют непрерывно.
9. Автоматизированный многомодульный инструмент для редактирования клеток по п. 6, в котором процессор выполнен с возможностью регулирования условий для выращивания в модуле для выращивания таким образом, что клетки достигают целевой оптической плотности в момент времени, запрошенный пользователем.
10. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, в котором емкость, выполненная с возможностью приема клеток, и одна или более емкостей, выполненных с возможностью приема нуклеиновых кислот, содержатся в картридже с реагентами.
11. Автоматизированный многомодульный инструмент для редактирования клеток по п. 10, в котором некоторые или все реагенты, необходимые для редактирования клеток, содержатся внутри картриджа с реагентами.
12. Автоматизированный многомодульный инструмент для редактирования клеток по п. 11, в котором реагенты, содержащиеся в картридже с реагентами, обнаруживают с помощью скрипта, считываемого процессором.
13. Автоматизированный многомодульный инструмент для редактирования клеток по п. 12, в котором картридж с реагентами включает реагенты и предоставляется в наборе.
14. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, в котором модуль трансформации содержит устройство электропорации.
15. Автоматизированный многомодульный инструмент для редактирования клеток по п. 14, в котором устройство электропорации представляет собой проточное устройство электропорации.
16. Автоматизированный многомодульный инструмент для редактирования клеток по п. 1, дополнительно содержащий модуль фильтрации, выполненный с возможностью концентрировать клетки и делать клетки электрокомпетентными.
17. Автоматизированный многомодульный инструмент для редактирования клеток, содержащий:
корпус, выполненный с возможностью размещения некоторых или всех модулей;
емкость, выполненную с возможностью приема клеток;
по меньшей мере, одну емкость, выполненную с возможностью приема нуклеиновых кислот;
модуль сборки нуклеиновых кислот, выполненный с возможностью сборки остова вектора и редактирующей кассеты, причем модуль сборки нуклеиновых кислот выполнен с возможностью приема и сборки нуклеиновых кислот для облегчения осуществления требуемого события редактирования генома в клетках;
модуль для выращивания, выполненный с возможностью выращивания клеток;
модуль трансформации, содержащий электропоратор для введения собранных нуклеиновых кислот в клетки;
модуль направляемого нуклеазой редактирования, выполненный с возможностью редактирования нуклеиновых кислот в клетках с помощью собранных нуклеиновых кислот;
автоматизированную систему подачи жидкости для перемещения жидкостей непосредственно из одного из: модуля сборки нуклеиновых кислот, модуля трансформации или модуля направляемого нуклеазой редактирования в другой из: модуля сборки нуклеиновых кислот, модуля трансформации или модуля направляемого нуклеазой редактирования без вмешательства пользователя; и
процессор, выполненный с возможностью обеспечения работы автоматизированного многомодульного инструмента для редактирования клеток, основываясь на вводе данных пользователем и/или выборе пользователем предварительно подготовленного скрипта.
18. Автоматизированный многомодульный инструмент для редактирования клеток по п. 17, дополнительно содержащий, по меньшей мере, один картридж с реагентами, содержащий реагенты, для выполнения редактирования клеток в автоматизированном многомодульном инструменте для редактирования клеток.
19. Автоматизированный многомодульный инструмент для редактирования клеток по п. 18, в котором емкости для клеток и нуклеиновых кислот расположены внутри картриджа с реагентами.
20. Автоматизированный многомодульный инструмент для редактирования клеток, содержащий:
корпус, выполненный с возможностью размещения некоторых или всех модулей;
емкость, выполненную с возможностью приема клеток;
по меньшей мере, одну емкость, выполненную с возможностью приема нуклеиновых кислот;
модуль сборки нуклеиновых кислот, выполненный с возможностью а) сборки остова и редактирующей кассеты и b) обессоливания собранных нуклеиновых кислот после сборки;
модуль для выращивания, выполненный с возможностью выращивания клеток;
модуль фильтрации, выполненный с возможностью концентрировать клетки и делать клетки электрокомпетентными;
модуль трансформации, содержащий проточный электропоратор для введения собранных нуклеиновых кислот в клетки;
комбинированный модуль восстановления и направляемого нуклеазой редактирования, выполненный с возможностью восстановления клеток после электропорации в модуле трансформации и редактирования клеток с помощью нуклеиновых кислот;
автоматизированную систему подачи жидкости для перемещения жидкостей непосредственно из одного из модуля для выращивания, модуля фильтрации, модуля трансформации, или комбинированного модуля восстановления и направляемого нуклеазой редактирования в другой из: модуля для выращивания, модуля фильтрации, модуля трансформации или комбинированного модуля восстановления и направляемого нуклеазой редактирования без вмешательства пользователя; и
процессор, выполненный с возможностью обеспечения работы автоматизированного многомодульного инструмента для редактирования клеток, основываясь на вводе данных пользователем.
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