Brief Communication | Published:

Efficient genome editing in zebrafish using a CRISPR-Cas system

Nature Biotechnology volume 31, pages 227229 (2013) | Download Citation

Abstract

In bacteria, foreign nucleic acids are silenced by clustered, regularly interspaced, short palindromic repeats (CRISPR)–CRISPR-associated (Cas) systems. Bacterial type II CRISPR systems have been adapted to create guide RNAs that direct site-specific DNA cleavage by the Cas9 endonuclease in cultured cells. Here we show that the CRISPR-Cas system functions in vivo to induce targeted genetic modifications in zebrafish embryos with efficiencies similar to those obtained using zinc finger nucleases and transcription activator–like effector nucleases.

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Acknowledgements

This work was supported by a US National Institutes of Health (NIH) Director's Pioneer Award DP1 GM105378 (J.K.J.), NIH R01 GM088040 (J.K.J. & R.T.P.), NIH P50 HG005550 (J.K.J.), the Jim and Ann Orr Research Scholar Award (J.K.J.), NIH K01 AG031300 (J.-R.J.Y.) and a Massachusetts General Hospital Claflin award (J.-R.J.Y.). We thank G. Church, J. Aach and P. Mali for sharing unpublished results and helpful discussions.

Author information

Author notes

    • Woong Y Hwang
    •  & Yanfang Fu

    These authors contributed equally to this work.

Affiliations

  1. Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.

    • Woong Y Hwang
    • , Randall T Peterson
    •  & J-R Joanna Yeh
  2. Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.

    • Yanfang Fu
    • , Deepak Reyon
    • , Morgan L Maeder
    • , Shengdar Q Tsai
    • , Jeffry D Sander
    •  & J Keith Joung
  3. Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.

    • Yanfang Fu
    • , Deepak Reyon
    • , Shengdar Q Tsai
    • , Jeffry D Sander
    •  & J Keith Joung
  4. Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, Massachusetts, USA.

    • Morgan L Maeder
    •  & J Keith Joung
  5. Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.

    • Randall T Peterson
    •  & J-R Joanna Yeh
  6. Broad Institute, Cambridge, Massachusetts, USA.

    • Randall T Peterson

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Contributions

W.Y.H., Y.F., M.L.M., D.R., S.Q.T., J.D.S., R.T.P., J.-R.J.Y. and J.K.J. designed experiments. W.Y.H., Y.F., M.L.M., D.R., S.Q.T. and J.D.S. performed experiments. W.Y.H., Y.F., M.L.M., D.R., S.Q.T., J.D.S., R.T.P., J.-R.J.Y. and J.K.J. wrote the paper.

Competing interests

J.K.J. has a financial interest in Transposagen Biopharmaceuticals.

Corresponding authors

Correspondence to J-R Joanna Yeh or J Keith Joung.

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    Supplementary Text and Figures

    Supplementary Discussion, Supplementary Methods, Supplementary Figures 1–5 and Supplementary Tables 1–5

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DOI

https://doi.org/10.1038/nbt.2501

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