Refined sgRNA efficacy prediction improves large- and small-scale CRISPR-Cas9 applications.

ABSTRACT: Genome editing with the CRISPR-Cas9 system has enabled unprecedented efficacy for reverse genetics and gene correction approaches. While off-target effects have been successfully tackled, the effort to eliminate variability in sgRNA efficacies-which affect experimental sensitivity-is in its infancy. To address this issue, studies have analyzed the molecular features of highly active sgRNAs, but independent cross-validation is lacking. Utilizing fluorescent reporter knock-out assays with verification at selected endogenous loci, we experimentally quantified the target efficacies of 430 sgRNAs. Based on this dataset we tested the predictive value of five recently-established prediction algorithms. Our analysis revealed a moderate correlation (r = 0.04 to r = 0.20) between the predicted and m

SUBMITTER: Labuhn M 

PROVIDER: S-EPMC5814880 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature