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Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins

Nature Biotechnology volume 34, pages 807–808 (2016)Cite this article

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Cpf1 represents a class 2/type V CRISPR RNA-guided endonuclease1 that is distinct from the type II CRISPR-Cas9 nuclease, widely used for genome editing2,3,4,5. It recognizes thymidine-rich protospacer-adjacent motif (PAM) sequences, expanding the range of RNA-guided genome editing beyond guanosine-rich PAM sequences recognized by Streptococcus pyogenes Cas9 (SpCas9). Yet, the potential of Cpf1 for targeted mutagenesis in whole organisms has not been demonstrated. Here, we report generation of mutant mice using preassembled recombinant CRISPR-Cpf1 ribonucleoproteins (RNPs).

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Figure 1: Cpf1 RNP-mediated genome editing in mouse embryos by electroporation.

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Acknowledgements

We thank Ji-hyun Ju for the help with the next-generation sequencing, and the members of the Genome Engineering Lab for helpful discussions and comments. This work was supported by the Institute for Basic Science (IBS-R021-D1 to J.-S.K.).

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Author notes

  1. Junho K Hur and Kyoungmi Kim: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Genome Engineering, Institute for Basic Science, Seoul, Republic of Korea

    Junho K Hur, Kyoungmi Kim, Kyung Wook Been, Gayoung Baek, Sunghyeok Ye, Junseok W Hur, Seuk-Min Ryu, Youn Su Lee & Jin-Soo Kim

  2. Department of Chemistry, Seoul National University, Seoul, Republic of Korea

    Kyung Wook Been, Seuk-Min Ryu & Jin-Soo Kim

  3. Basic Science, IBS School, Korea University of Science and Technology, Seoul, Republic of Korea

    Sunghyeok Ye

  4. Department of Neurosurgery, Korea University College of Medicine, Seoul, Republic of Korea

    Junseok W Hur

  5. ToolGen, Seoul, Republic of Korea

    Youn Su Lee

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  1. Junho K Hur
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Correspondence to Jin-Soo Kim.

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J.-S.K., J.K.H., and K.M.K. have filed a patent application based on this work.

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Editor's note: This article has been peer reviewed.

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Hur, J., Kim, K., Been, K. et al. Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins. Nat Biotechnol 34, 807–808 (2016). https://doi.org/10.1038/nbt.3596

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