Genomic editing in automated systems
AssigneeInscripta, Inc.
InventorsChristopher Abraham, Charles Johnson, Clint Davis, Eileen Spindler, Paul Hardenbol
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Abstract
The present disclosure thus provides methods for higher efficiency cell editing that maintain the viability of the cells. In certain aspects, the present disclosure provides systems and instruments for automated methods of multiplexed nuclease-directed genome editing using nuclease expression under a constitutive promoter. In specific aspects, the constitutive promoter is an insulated promoter. In other aspects, the constitutive promoter is a heterologous promoter. In still other aspects, the constitutive promoter is a synthetic promoter.
Claims17
A method of modifying a target region in the genome of a cell, the method comprising:
- (a) contacting a cell with:
- a nucleic-acid-guided nuclease encoded by a nucleic acid operably linked to a heterologous constitutive promoter;
- a guide nucleic acid capable of complexing with the nucleic acid-guided nuclease;
- and
- an editing sequence encoding a nucleic acid complementary to said target region having a change in sequence relative to the target region;
- and
- (b) allowing the nuclease, guide nucleic acid, and editing sequence to create a genome edit in a target region of the genome of the cell.
A composition for use in genome editing comprising a nuclease operably linked to a heterologous constitutive promoter optimized for use in cells from a particular organism.
An automated system for multiplexed nuclease-directed genome editing, comprising:
- a nuclease operably linked to a heterologous constitutive promoter;
- and
- an instrument comprising:
- a housing,
- one or more receptacles to receive cells and one or more nucleic acids comprising sequences to facilitate nuclease-directed genome editing in the cells;
- a unit for introduction of the nucleic acid(s) into the cells;
- a unit for allowing the nuclease-directed genome editing events to occur;
- and
- a unit for configuring the operation of the system based on user input.
Dependencies are inferred from the claim text.