Journal article
Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro
Biochemistry (Easton), Vol.54(48), pp.7063-7066
12/08/2015
Handle:
https://hdl.handle.net/2376/116233
PMID: 26579937
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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Details
- Title
- Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro
- Creators
- John M Hinz - School of Molecular Biosciences and Center for Reproductive Biology, Washington State University , Pullman, Washington 99164-7520, United StatesMarian F Laughery - School of Molecular Biosciences and Center for Reproductive Biology, Washington State University , Pullman, Washington 99164-7520, United StatesJohn J Wyrick - School of Molecular Biosciences and Center for Reproductive Biology, Washington State University , Pullman, Washington 99164-7520, United States
- Publication Details
- Biochemistry (Easton), Vol.54(48), pp.7063-7066
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- United States
- Grant note
- ES004106 / NIEHS NIH HHS ES002614 / NIEHS NIH HHS
- Identifiers
- 99900548069001842
- Language
- English
- Resource Type
- Journal article