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Generation of knockout mice by Cpf1-mediated gene targeting

Nature Biotechnology volume 34, pages 808–810 (2016)Cite this article

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To the Editor:

Engineered nucleases, including transcription activator-like effector nucleases (TALENs) and CRISPR-Cas9, are under active development as versatile genome-editing tools in various organisms including mice1,2,3. Recently, diverse Cpf1 orthologs were identified as representatives of a class 2 CRISPR-Cas system in vitro and showed target-specific endonuclease activities in human cells comparable to that of Streptococcus pyogenes Cas9 (SpCas9)4. In contrast to Cas9, Cpf1 is guided by a single CRISPR RNA (crRNA) without an additional trans-activating crRNA, requires a short thymidine-rich protospacer-adjacent motif (PAM) preceding the target sequence, and introduces a staggered double-stranded break4. These simple characteristics of Cpf1 should be of great use for the generation of knockout mice.

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Figure 1: Generation of Cpf1-mediated Trp53 mutant mice.

References

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Acknowledgements

This study was supported by the Ministry of Science, ICT and Future Planning (MSIP) through the National Research Foundation of Korea (NRF) (2013R1A1A1009962, 2014M3A9D5A01073836, 2015R1A1A1A05001483). This work was also supported by a grant (2016-649) from the Asan Institute for Life Sciences, Asan Medical Center, Seoul, Korea. We appreciate the technical assistance from the Genomics Core Laboratory of Asan Institute for Life Sciences.

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Authors and Affiliations

  1. Department of Biomedical Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea

    Yongsub Kim & Myeong Sup Lee

  2. Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea

    Seung-A Cheong, Jong Geol Lee, In-Jeoung Baek & Young Hoon Sung

  3. College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea

    Jong Geol Lee

  4. Department of Radiation Oncology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea

    Sang-Wook Lee

  5. Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul, Republic of Korea

    In-Jeoung Baek & Young Hoon Sung

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  1. Yongsub Kim
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  2. Seung-A Cheong
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Correspondence to Sang-Wook Lee, Myeong Sup Lee, In-Jeoung Baek or Young Hoon Sung.

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Supplementary Figures 1–10, Supplementary Tables 1–5 and Supplementary Methods (PDF 2686 kb)

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Kim, Y., Cheong, SA., Lee, J. et al. Generation of knockout mice by Cpf1-mediated gene targeting. Nat Biotechnol 34, 808–810 (2016). https://doi.org/10.1038/nbt.3614

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