Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo

Published on Sep 18, 2018in Proceedings of the National Academy of Sciences of the United States of America9.58
· DOI :10.1073/pnas.1810062115
Robert M. Yarrington7
Estimated H-index: 7
(UofU: University of Utah),
Surbhi Verma6
Estimated H-index: 6
(UofU: University of Utah)
+ 2 AuthorsDana Carroll43
Estimated H-index: 43
(UofU: University of Utah)
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 is relieved when nucleosomes are depleted. Remarkably, the same is not true of zinc-finger nucleases (ZFNs), which cleave equally well at nucleosome-occupied and nucleosome-depleted sites. These results have implications for the choice of specific targets for genome editing, both in research and in clinical and other practical applications.
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