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Efficient genome editing in zebrafish using a CRISPR-Cas system

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

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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 We thank G. Church, J. Aach and P. Mali for sharing unpublished results and helpful discussions.

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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.

Corresponding authors

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

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Competing interests

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

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

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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

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