Francisella novicida Cas9 interrogates genomic DNA with very high specificity and can be used for mammalian genome editing.

ABSTRACT: Genome editing using the CRISPR/Cas9 system has been used to make precise heritable changes in the DNA of organisms. Although the widely used Streptococcus pyogenes Cas9 (SpCas9) and its engineered variants have been efficiently harnessed for numerous gene-editing applications across different platforms, concerns remain regarding their putative off-targeting at multiple loci across the genome. Here we report that Francisella novicida Cas9 (FnCas9) shows a very high specificity of binding to its intended targets and negligible binding to off-target loci. The specificity is determined by its minimal binding affinity with DNA when mismatches to the target single-guide RNA (sgRNA) are present in the sgRNA:DNA heteroduplex. FnCas9 produces staggered cleavage, higher homology-direc

SUBMITTER: Acharya S 

PROVIDER: S-EPMC6800334 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature