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Heritable multiplex genetic engineering in rats using CRISPR/Cas9.

ABSTRACT: The CRISPR/Cas9 system has been proven to be an efficient gene-editing tool for genome modification of cells and organisms. Multiplex genetic engineering in rat holds a bright future for the study of complex disease. Here, we show that this system enables the simultaneous disruption of four genes (ApoE, B2m, Prf1, and Prkdc) in rats in one-step, by co-injection of Cas9 mRNA and sgRNAs into fertilized eggs. We further observed the gene modifications are germline transmittable, and confirmed the off-target mutagenesis and mosaicism are rarely detected by comprehensive analysis. Thus, the CRISPR/Cas9 system makes it possible to efficiently and reliably generate gene knock-out rats.

SUBMITTER: Ma Y

PROVIDER: S-EPMC3943732 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Heritable multiplex genetic engineering in rats using CRISPR/Cas9.

Ma Yuanwu Y Shen Bin B Zhang Xu X Lu Yingdong Y Chen Wei W Ma Jing J Huang Xingxu X Zhang Lianfeng L

PloS one 20140305 3

The CRISPR/Cas9 system has been proven to be an efficient gene-editing tool for genome modification of cells and organisms. Multiplex genetic engineering in rat holds a bright future for the study of complex disease. Here, we show that this system enables the simultaneous disruption of four genes (ApoE, B2m, Prf1, and Prkdc) in rats in one-step, by co-injection of Cas9 mRNA and sgRNAs into fertilized eggs. We further observed the gene modifications are germline transmittable, and confirmed the o ...[more]

PMID: 24598943

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