Journal article
Nucleosomes Selectively Inhibit Cas9 Off-target Activity at a Site Located at the Nucleosome Edge
The Journal of biological chemistry, Vol.291(48), pp.24851-24856
11/25/2016
Handle:
https://hdl.handle.net/2376/116224
PMCID: PMC5122757
PMID: 27756838
Abstract
Nucleosomes affect Cas9 binding and activity at on-target sites, but their impact at off-target sites is unknown. To investigate how nucleosomes affect Cas9 cleavage at off-target sites in vitro, we used a single guide RNA (sgRNA) that has been previously shown to efficiently direct Cas9 cleavage at the edge of the strongly positioned 601 nucleosome. Our data indicate that single mismatches between the sgRNA and DNA target have relatively little effect on Cas9 cleavage of naked DNA substrates, but strongly inhibit cleavage of nucleosome substrates, particularly when the mismatch is in the sgRNA "seed" region. These findings indicate that nucleosomes may enhance Cas9 specificity by inhibiting cleavage of off-target sites at the nucleosome edge.
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Details
- Title
- Nucleosomes Selectively Inhibit Cas9 Off-target Activity at a Site Located at the Nucleosome Edge
- Creators
- John M Hinz - From the School of Molecular Biosciences and Center for Reproductive Biology, Washington State University, Pullman, Washington 99164Marian F Laughery - From the School of Molecular Biosciences and Center for Reproductive Biology, Washington State University, Pullman, Washington 99164John J Wyrick - From the School of Molecular Biosciences and Center for Reproductive Biology, Washington State University, Pullman, Washington 99164 jwyrick@vetmed.wsu.edu
- Publication Details
- The Journal of biological chemistry, Vol.291(48), pp.24851-24856
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- United States
- Grant note
- R37 ES002614 / NIEHS NIH HHS R01 ES002614 / NIEHS NIH HHS
- Identifiers
- 99900548267201842
- Language
- English
- Resource Type
- Journal article