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Nat Methods. 2018 Jul;15(7):512-514. doi: 10.1038/s41592-018-0011-5. Epub 2018 May 21.

CRISPR off-target analysis in genetically engineered rats and mice.

Author information

1
Department of Molecular Biology, Genentech, Inc., South San Francisco, CA, USA.
2
Jack Baskin School of Engineering, University of California, Santa Cruz, Santa Cruz, CA, USA.
3
Bioinformatics & Computational Biology, Genentech, Inc., South San Francisco, CA, USA.
4
Laboratory Animal Resources, Genentech, Inc., South San Francisco, CA, USA.
5
Department of Molecular Biology, Genentech, Inc., South San Francisco, CA, USA. sorenw@gene.com.

Abstract

Despite widespread use of CRISPR, comprehensive data on the frequency and impact of Cas9-mediated off-targets in modified rodents are limited. Here we present deep-sequencing data from 81 genome-editing projects on mouse and rat genomes at 1,423 predicted off-target sites, 32 of which were confirmed, and show that high-fidelity Cas9 versions reduced off-target mutation rates in vivo. Using whole-genome sequencing data from ten mouse embryos, treated with a single guide RNA (sgRNA), and from their genetic parents, we found 43 off-targets, 30 of which were predicted by an adapted version of GUIDE-seq.

PMID:
29786090
PMCID:
PMC6558654
DOI:
10.1038/s41592-018-0011-5
[Indexed for MEDLINE]
Free PMC Article

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