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Genome-scale loss-of-function screening with a lentiviral RNAi library

Nature Methods volume 3, pages 715–719 (2006)Cite this article

Abstract

The discovery that RNA interference (RNAi) is functional in mammalian cells led us to form The RNAi Consortium (TRC) with the goal of enabling large-scale loss-of-function screens through the development of genome-scale RNAi libraries and methodologies for their use. These resources form the basis of a loss-of-function screening platform created at the Broad Institute. Our human and mouse libraries currently contain >135,000 lentiviral clones targeting 27,000 genes. Initial screening efforts have demonstrated that these libraries and methods are practical and powerful tools for high-throughput lentivirus RNAi screens.

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Figure 1: Vector map for the pLKO.1 lentiviral vector.

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Acknowledgements

The authors thank J. Grenier, G. Hinkle, B. Luo, J. Moffat and S. Silver for their contributions to the work reviewed here and for their comments on the manuscript. The TRC library is the product of The RNAi Consortium (TRC). We thank the members of TRC (Academia Sinica, Bristol-Meyers Squibb, Eli Lilly, Novartis, Sigma-Aldrich, the Broad Institute of MIT and Harvard, Dana-Farber Cancer Institute, Massachusetts General Hospital, Harvard Medical School, the Massachusetts Institute of Technology, and the Whitehead Institute for Biomedical Research) for their scientific contributions and financial support. The Broad Institute and TRC do not profit from the distribution of the TRC library. Sigma-Aldrich, one of five sponsoring members of The RNAi Consortium who contributed equally to funding the work described here, is a commercial distributor of TRC shRNA library clones.

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Authors and Affiliations

  1. Broad Institute of MIT and Harvard, Cambridge, 02142, Massachusetts, USA

    David E Root, Nir Hacohen, William C Hahn, Eric S Lander & David M Sabatini

  2. Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Nir Hacohen

  3. Harvard Medical School, Boston, 02115, Massachusetts, USA

    Nir Hacohen, William C Hahn & Eric S Lander

  4. Department of Medical Oncology, Center for Cancer Systems Biology, Center for Cancer Genome Discovery, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    William C Hahn

  5. Whitehead Institute for Biomedical Research, Cambridge, 02142, Massachusetts, USA

    Eric S Lander & David M Sabatini

  6. Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Eric S Lander & David M Sabatini

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  1. David E Root
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  2. Nir Hacohen
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Correspondence to David E Root.

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Root, D., Hacohen, N., Hahn, W. et al. Genome-scale loss-of-function screening with a lentiviral RNAi library. Nat Methods 3, 715–719 (2006). https://doi.org/10.1038/nmeth924

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