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Heritable gene targeting in the mouse and rat using a CRISPR-Cas system

Nature Biotechnology volume 31, pages 681–683 (2013)Cite this article

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CRISPR-Cas systems have been developed as an efficient gene editing technology in cells and model organisms. Here we use a CRISPR-Cas system to induce genomic DNA fragment deletion in mice by co-injecting two single-guide RNAs (sgRNAs) targeting the Uhrf2 locus with Cas9 mRNA. Furthermore, we report the generation of a Mc3R and Mc4R double-gene knockout rat by means of a single microinjection. High germline-transmission efficiency was observed in both mice and rats.

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Figure 1: Generation of gene mutant rats with a CRISPR-Cas system.

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Acknowledgements

We thank F. Zhang of the Broad Institute of MIT and Harvard for kindly providing us with the Cas9 expression vector. We thank S. Siwko for comments and advice. We also thank S.S. Bae and J.-S. Kim of Seoul National University for helping us to predict the potential off-target sites. This work was partially supported by grants from the State Key Development Programs of China (2012CB910400 to M.L., 2010CB945403 to D.L.), grants from the National Natural Science Foundation of China (31171318 to D.L. and 30930055 to M.L.), a grant from the Science and Technology Commission of Shanghai Municipality (11DZ2260300) and grants from the Program for Changjiang Scholars and Innovative Research Team in University (IRT1119 and IRT1128).

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Author notes

  1. Dali Li and Zhongwei Qiu: These authors contributed equally to this work.

  2. myliu@bio.ecnu.edu.cn

  3. yongxiang_zhao@126.com

Authors and Affiliations

  1. Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China

    Dali Li, Zhongwei Qiu, Yanjiao Shao, Yuting Chen, Yuting Guan, Meizhen Liu, Yongmei Li, Na Gao, Liren Wang & Mingyao Liu

  2. Biological Targeting Diagnosis and Therapy Research Center, Guangxi Medical University, Nanning, Guangxi, China

    Xiaoling Lu & Yongxiang Zhao

  3. The Institute of Biosciences and Technology, Texas A&M University Health Science Center, Houston, Texas, USA

    Mingyao Liu

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  1. Dali Li
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Correspondence to Dali Li, Yongxiang Zhao or Mingyao Liu.

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Supplementary Methods, Supplementary Figures 1–9 and Supplementary Table 1 (PDF 936 kb)

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Li, D., Qiu, Z., Shao, Y. et al. Heritable gene targeting in the mouse and rat using a CRISPR-Cas system. Nat Biotechnol 31, 681–683 (2013). https://doi.org/10.1038/nbt.2661

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