Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo.

ABSTRACT: Genome editing with CRISPR-Cas nucleases has been applied successfully to a wide range of cells and organisms. There is, however, considerable variation in the efficiency of cleavage and outcomes at different genomic targets, even within the same cell type. Some of this variability is likely due to the inherent quality of the interaction between the guide RNA and the target sequence, but some may also reflect the relative accessibility of the target. We investigated the influence of chromatin structure, particularly the presence or absence of nucleosomes, on cleavage by the Streptococcus pyogenes Cas9 protein. At multiple target sequences in two promoters in the yeast genome, we find that Cas9 cleavage is strongly inhibited when the DNA target is within a nucleosome. This inhibition

SUBMITTER: Yarrington RM 

PROVIDER: S-EPMC6156633 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature