DNA targeting specificity of RNA-guided Cas9 nucleases

Analyses of the determinants of the specificity of Cas9 nuclease provide rules for selecting optimal target sites. The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by means of single-guide RNAs (sgRNAs) to enable genome editing1,2,3,4,5,6,7,8,9,10. Here, we characterize SpCas9 targeting specificity in human cells to inform the selection of target sites and avoid off-target effects. Our study evaluates...