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Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring.


ABSTRACT: The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. Here, we analyzed whole-genome sequencing (WGS) data from two knock-out (KO) mouse strains generated by using the CRISPR-Cas9 system targeting the Mmd and Paqr8 loci. A total of nine individuals were sequenced including two parents, four F1 offspring and three uninjected control mice. Using GATK and bcftools software, we identified two off-target events in the founder mice. The two CRISPR-Cas9-induced off-target events were predictable using Cas-OFFinder and were not passed on to the offspring that we investigated. In addition, our results indicated that the number of CRISPR-Cas9-induced mutations was not statistically distinguishable from the background de novo mutations (DNMs).

SUBMITTER: Dong Y 

PROVIDER: S-EPMC6829146 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring.

Dong Yan Y   Li Haimei H   Zhao Liang L   Koopman Peter P   Zhang Feng F   Huang Johnny X JX  

G3 (Bethesda, Md.) 20191105 11


The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. Here, we analyzed whole-genome sequencing (WGS) data fro  ...[more]

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