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Improved vectors and genome-wide libraries for CRISPR screening

Nature Methods volume 11, pages 783–784 (2014)Cite this article

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Genome-wide, targeted loss-of-function pooled screens using the clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated nuclease Cas9 in human and mouse cells provide an alternative screening system to RNA interference (RNAi)1,2,3,4. Previously, we used a genome-scale CRISPR knockout (GeCKO) library to identify loss-of-function mutations conferring vemurafenib resistance in a melanoma model1. However, initial lentiviral delivery systems for CRISPR screening had low viral titer or required a cell line already expressing Cas9, thereby limiting the range of biological systems amenable to screening.

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Figure 1: CRISPR lentiviral vectors with higher functional titer.

References

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Acknowledgements

We thank the Zhang lab for support and advice. N.E.S. is supported by a postdoctoral fellowship from the Simons Center for the Social Brain at Massachusetts Institute of Technology. O.S. is supported by a postdoctoral fellowship from the Klarman Cell Observatory. F.Z. is supported by the US National Institutes of Health and NIMH through a Director's Pioneer Award (5DP1-MH100706); a Transformative R01 grant (5R01-DK097768); the Keck, Merkin, Vallee, Damon Runyon, Searle Scholars, Klingenstein and Simons Foundations; and by B. Metcalfe and J. Pauley.

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Author notes

  1. Neville E Sanjana and Ophir Shalem: These authors contributed equally to this work.

Authors and Affiliations

  1. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Neville E Sanjana, Ophir Shalem & Feng Zhang

  2. McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Neville E Sanjana, Ophir Shalem & Feng Zhang

  3. Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Neville E Sanjana, Ophir Shalem & Feng Zhang

  4. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Neville E Sanjana, Ophir Shalem & Feng Zhang

Authors
  1. Neville E Sanjana
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  2. Ophir Shalem
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  3. Feng Zhang
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Contributions

N.E.S., O.S. and F.Z. conceived of and designed the experiments. N.E.S. and O.S. performed the experiments and analyzed the data. N.S., O.S. and F.Z. wrote the manuscript.

Corresponding author

Correspondence to Feng Zhang.

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Competing interests

N.E.S., O.S. and F.Z. are on patent applications related to this work. F.Z. is a cofounder of and scientific advisor for Editas Medicine.

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Supplementary Figures 1–3, Supplementary Methods and Supplementary Data (PDF 638 kb)

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Sanjana, N., Shalem, O. & Zhang, F. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods 11, 783–784 (2014). https://doi.org/10.1038/nmeth.3047

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