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US 11,739,290 B2Granted
Instruments, modules, and methods for improved detection of edited sequences in live cells
AssigneeInscripta, Inc.
InventorsAndrew Garst, Richard Fox, Phillip Belgrader, Don Masquelier
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Abstract
The present disclosure provides instruments, modules and methods for improved detection of edited cells following nucleic acid-guided nuclease genome editing. The disclosure provides improved automated instruments that perform methods—including high throughput methods—for screening cells that have been subjected to editing and identifying cells that have been properly edited.
Claims20
A method for enriching edited cells comprising:
- transforming cells with one or more vectors comprising a first promoter driving transcription of a coding sequence for a CRISPR nuclease and a second promoter driving transcription of a guide nucleic acid sequence;
- diluting the transformed cells to a cell concentration to substantially cingulate the transformed cells;
- growing the substantially singulated transformed cells;
- initiating editing by inducing the second promoter for driving transcription of the guide nucleic acid sequence;
- growing the edited cells to form colonies;
- and
- selecting or pooling slow-growing colonies, of the colonies, wherein cells from the slow-growing colonies are enriched for edited cells.
4 more elements
A method for enriching edited comprising:
- transforming cells with one or more vectors comprising a first promoter driving transcription of a coding sequence for a CRISPR nuclease and a second promoter driving transcription of a guide nucleic acid sequence;
- diluting the transformed cells to a cell concentration to substantially cingulate the transformed cells;
- growing the substantially singulated transformed cells until the substantially singulated transformed cells establish colonies having respective colony sizes;
- initiating editing by inducing the second promoter;
- growing the edited cells to form colonies of terminal size;
- and
- harvesting the colonies of terminal size.
4 more elements
Dependencies are inferred from the claim text.