Heritable multiplex genetic engineering in rats using CRISPR/Cas9.

Ma, Yuanwu; Shen, Bin; Zhang, Xu; Lu, Yingdong; Chen, Wei; Ma, Jing; Huang, Xingxu; Zhang, Lianfeng · PLoS One · 2014

basic_science · Level V

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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.

Medical subject headings