Skip to main content
SpringerLink
Log in

Efficient genome editing in zebrafish using a CRISPR-Cas system

  • Brief Communication
  • Published:

From Nature Biotechnology

View current issue Submit your manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1: Schematic illustrating naturally occurring and engineered CRISPR-Cas systems.

Similar content being viewed by others

References

  1. Wiedenheft, B., Sternberg, S.H. & Doudna, J.A. Nature 482, 331–338 (2012).

    Article  CAS  Google Scholar 

  2. Horvath, P. & Barrangou, R. Science 327, 167–170 (2010).

    Article  CAS  Google Scholar 

  3. Terns, M.P. & Terns, R.M. Curr. Opin. Microbiol. 14, 321–327 (2011).

    Article  CAS  Google Scholar 

  4. Barrangou, R. et al. Science 315, 1709–1712 (2007).

    Article  CAS  Google Scholar 

  5. Brouns, S.J. et al. Science 321, 960–964 (2008).

    Article  CAS  Google Scholar 

  6. Jinek, M. et al. Science 337, 816–821 (2012).

    Article  CAS  Google Scholar 

  7. Gasiunas, G., Barrangou, R., Horvath, P. & Siksnys, V. Proc. Natl. Acad. Sci. USA 109, E2579–E2586 (2012).

    Article  CAS  Google Scholar 

  8. Mali, P. et al. Science doi:10.1126/science.1232033 (3 January 2013).

  9. Cong, L. et al. Science doi:10.1126/science.1231143 (3 January 2013).

  10. Urnov, F.D., Rebar, E.J., Holmes, M.C., Zhang, H.S. & Gregory, P.D. Nat. Rev. Genet. 11, 636–646 (2010).

    Article  CAS  Google Scholar 

  11. Joung, J.K. & Sander, J.D. Nat. Rev. Mol. Cell Biol. 14, 49–55 (2012).

    Article  Google Scholar 

  12. Holkers, M. et al. Nucleic Acids Res. doi:10.1093/nar/gks1446 (28 December 2012).

    Article  CAS  Google Scholar 

  13. Schneider, T.D. & Stormo, G.D. Nucleic Acids Res. 17, 659–674 (1989).

    Article  CAS  Google Scholar 

  14. Sander, J.D. et al. Nucleic Acids Res. 38, W462–W468 (2010).

    Article  CAS  Google Scholar 

  15. Foley, J.E. et al. PLoS ONE 4, e4348 (2009).

    Article  Google Scholar 

  16. Huang, P. et al. Nat. Biotechnol. 29, 699–700 (2011).

    Article  Google Scholar 

  17. Cade, L. et al. Nucleic Acids Res. 40, 8001–8010 (2012).

    Article  CAS  Google Scholar 

  18. Sander, J.D. et al. Nat. Biotechnol. 29, 697–698 (2011).

    Article  CAS  Google Scholar 

  19. Sander, J.D. et al. Nat. Methods 8, 67–69 (2011).

    Article  CAS  Google Scholar 

  20. Zuker, M. Nucleic Acids Res. 31, 3406–3415 (2003).

    Article  CAS  Google Scholar 

Download references

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

  1. Woong Y Hwang and Yanfang Fu: These authors contributed equally to this work.

Authors and 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

Authors
  1. Woong Y Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanfang Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Deepak Reyon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Morgan L Maeder
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shengdar Q Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jeffry D Sander
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Randall T Peterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J-R Joanna Yeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. J Keith Joung
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Corresponding authors

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

Ethics declarations

Competing interests

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

Supplementary information

Supplementary Text and Figures

Supplementary Discussion, Supplementary Methods, Supplementary Figures 1–5 and Supplementary Tables 1–5 (PDF 487 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hwang, W., Fu, Y., Reyon, D. et al. Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol 31, 227–229 (2013). https://doi.org/10.1038/nbt.2501

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

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

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation