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RNAi screens in Caenorhabditis elegans in a 96-well liquid format and their application to the systematic identification of genetic interactions

Nature Protocols volume 1, pages 1617–1620 (2006)Cite this article

Abstract

We describe a protocol for performing RNA interference (RNAi) screens in Caenorhabditis elegans in liquid culture in 96-well plates. The procedure allows a single researcher to set-up and score RNAi experiments at ∼2,000 genes per day. By comparing RNAi phenotypes between wild-type worms and worms carrying a defined genetic mutation, we have used this protocol to identify synthetic lethal interactions between genes systematically. We also describe how the protocol can be adapted to target two genes simultaneously by combinatorial RNAi.

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Figure 1
Figure 2: Example phenotypes.

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Acknowledgements

We thank the C. elegans Genetics Center for providing strains. B.L. was supported by a Sanger Institute Postdoctoral Fellowship. J.T., A.F. and A.G.F. were supported by the Wellcome Trust.

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Authors and Affiliations

  1. The Wellcome Trust Sanger Institute, Hinxton, CB10 1HH, Cambridge, UK

    Ben Lehner, Julia Tischler & Andrew G Fraser

  2. CRG-EMBL Systems Biology Program, Center for Genomic Regulation, c/Dr. Aiguader 88, Barcelona, 08003, Spain

    Ben Lehner

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  1. Ben Lehner
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  2. Julia Tischler
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  3. Andrew G Fraser
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Correspondence to Andrew G Fraser.

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Lehner, B., Tischler, J. & Fraser, A. RNAi screens in Caenorhabditis elegans in a 96-well liquid format and their application to the systematic identification of genetic interactions. Nat Protoc 1, 1617–1620 (2006). https://doi.org/10.1038/nprot.2006.245

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