Optimizing CRISPR/Cas9 Editing of Repetitive Single Nucleotide Variants.

ABSTRACT: CRISPR/Cas9, base editors and prime editors comprise the contemporary genome editing toolbox. Many studies have optimized the use of CRISPR/Cas9, as the original CRISPR genome editing system, in substituting single nucleotides by homology directed repair (HDR), although this remains challenging. Studies describing modifications that improve editing efficiency fall short of isolating clonal cell lines or have not been validated for challenging loci or cell models. We present data from 95 transfections using a colony forming and an immortalized cell line comparing the effect on editing efficiency of donor template modifications, concentration of components, HDR enhancing agents and cold shock. We found that in silico predictions of guide RNA efficiency correlated poorly withactivity i

SUBMITTER: Usher I 

PROVIDER: S-EPMC9294353 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature