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shRNA libraries and their use in cancer genetics

Nature Methods volume 3, pages 701–706 (2006)Cite this article

Abstract

RNA interference was originally described as a powerful tool to inhibit gene expression in model organisms. Until recently, loss-of-function genetic screens in mammalian cells were hampered by a lack of suitable tools that can be used in a high-throughput format. Here we discuss the construction of short-hairpin RNA (shRNA) vector libraries, in particular those generated at the Netherlands Cancer Institute (NKI), and their application in mammalian cancer genetics. We describe their virtues and limitations, as well as different options for screening such libraries.

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Figure 1: Construction of the NKI shRNA library.
Figure 2: siRNA bar-code screens.

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Acknowledgements

We thank A. Fabius and A. Dirac for critical reading of the manuscript. This work was supported by grants from the Netherlands Genomics Initiative, The EU 6th framework integrated project “INTACT”, the centre for Biomedical genetics (CBG) and the Dutch Cancer Society (KWF).

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Author notes

  1. Thijn R Brummelkamp

    Present address: Whitehead Institute, 9 Cambridge Center, Cambridge, Massachusetts, 02142, USA

Authors and Affiliations

  1. Division of Molecular Carcinogenesis and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    René Bernards, Thijn R Brummelkamp & Roderick L Beijersbergen

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  1. René Bernards
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  3. Roderick L Beijersbergen
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Correspondence to René Bernards or Roderick L Beijersbergen.

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R.B. and R.L.B. receive royalties on sales of the shRNA library.

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Bernards, R., Brummelkamp, T. & Beijersbergen, R. shRNA libraries and their use in cancer genetics. Nat Methods 3, 701–706 (2006). https://doi.org/10.1038/nmeth921

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