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US 12,612,662 B2Granted
DNA repair site detection for personal genomics, epigenomics, and gene therapy
AssigneeThe Salk Institute for Biological Studies
InventorsFred H. Gage, Dylan A. Reid, Patrick J. Reed
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Abstract
Provided are methods for identification of DNA repair locations in a genome of a non-dividing cell, by incorporating a reactive nucleoside analogs into the genome of the non-dividing cell, then sequencing the regions of the genome that incorporated the nucleoside analog.
Claims20
A method of identifying DNA repair locations in a genome of a non-dividing cell, comprising:
- incubating a non-dividing cell comprising genomic DNA (gDNA) with at least one reactive nucleoside analog under conditions that permit the at least one reactive nucleoside analog to be incorporated into the gDNA via a repair DNA polymerase of the non-dividing cell, thereby generating gDNA comprising the at least one reactive nucleoside analog;
- obtaining gDNA comprising the at least one reactive nucleoside analog from the non-dividing cells;
- generating a population of gDNA fragments, wherein the population of gDNA fragments comprises gDNA fragments comprising the at least one reactive nucleoside analog, and gDNA fragments that do not comprise the at least one reactive nucleoside analog;
- isolating from the population of gDNA fragments those gDNA fragments comprising the at least one reactive nucleoside analog;
- and
- sequencing the isolated gDNA fragments comprising the at least one reactive nucleoside analog, thereby identifying DNA repair locations in the genome of the non-dividing cell.
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Dependencies are inferred from the claim text.