Cell-free test method for measuring base editing ability of base editor
AssigneeEdgene, Inc.
InventorsJin Soo KIM, Seong Hyun LEE
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Abstract
The present invention relates to a method for testing genome editing in a cell-free, in vitro system and, in particular, to a method for identifying and quantifying target DNA mutations by a base editor without using eukaryotic cells and without separate complicated experimental procedures. By using the present invention, target DNA mutations can be identified and quantified by a base editor without using cells and without separate complicated experimental procedures. In addition, a base editor having the ability to edit target DNA mutations in actual cells can be screened through a simple cell-free test method.
Claims20
A method for providing information on the ability of a base editing system to correct a DNA mutation, the method comprising:
- (a) contacting an isolated DNA comprising a target DNA mutation with one or more base-editing proteins used in a base-editing system in a cell-free environment;
- (b) sequencing the DNA obtained from step (a) to measure the base-editing activity of the base-editing system;
- and
- (c) providing information on the ability of the base-editing system to correct the same DNA mutation in a cell.
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A method for screening a base editor capable of correcting a target DNA mutation in cells, the method comprising:
- (a) a step of contacting an isolated DNA comprising a target DNA mutation with candidate base editors in a cell-free environment, wherein the candidate base editors each independently comprise one or more base-editing proteins or one or more polynucleotides encoding the same;
- (b) measuring the base editing activity of the candidate base editors by sequencing the DNA molecules obtained from step (a);
- (c) providing information on the ability of the candidate base editors to correct the same DNA mutation in cells;
- and
- (d) selecting a base editor capable of correcting the same DNA mutation in cells based on the information obtained from step (c).
Dependencies are inferred from the claim text.