Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro - PubMed (original) (raw)

. 2015 Dec 8;54(48):7063-6.

doi: 10.1021/acs.biochem.5b01108. Epub 2015 Nov 24.

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Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro

John M Hinz et al. Biochemistry. 2015.

Abstract

During Cas9 genome editing in eukaryotic cells, the bacterial Cas9 enzyme cleaves DNA targets within chromatin. To understand how chromatin affects Cas9 targeting, we characterized Cas9 activity on nucleosome substrates in vitro. We find that Cas9 endonuclease activity is strongly inhibited when its target site is located within the nucleosome core. In contrast, the nucleosome structure does not affect Cas9 activity at a target site within the adjacent linker DNA. Analysis of target sites that partially overlap with the nucleosome edge indicates that the accessibility of the protospacer-adjacent motif (PAM) is the critical determinant of Cas9 activity on a nucleosome.

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