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Cas9 as a versatile tool for engineering biology

Nature Methods volume 10, pages 957–963 (2013)Cite this article

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Abstract

RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein, will grant unprecedented control over cellular organization, regulation and behavior. Here we describe the Cas9 targeting methodology, detail current and prospective engineering advances and suggest potential applications ranging from basic science to the clinic.

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Figure 1: Functioning of the type II CRISPR-Cas systems in bacteria3.
Figure 2: Cas9-sgRNA targeting complexes.
Figure 3: Platform for multiplex biological screens.
Figure 4: Cas9 therapeutics.

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Acknowledgements

We thank J. Aach and L. Yang for insightful discussions, and W.L. Chew for insightful discussions and critical reading of the manuscript. This work was supported by US National Institutes of Health NHGRI grant P50 HG005550, Department of Energy grant DE-FG02-02ER63445 and the Wyss Institute for Biologically Inspired Engineering (K.M.E.).

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  1. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Prashant Mali & George M Church

  2. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA

    Kevin M Esvelt & George M Church

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  1. Prashant Mali
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Correspondence to Prashant Mali or Kevin M Esvelt.

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Mali, P., Esvelt, K. & Church, G. Cas9 as a versatile tool for engineering biology. Nat Methods 10, 957–963 (2013). https://doi.org/10.1038/nmeth.2649

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