RNAi screening to identify postembryonic phenotypes in C. elegans.

Journal of Visualized Experiments : JoVE
Katherine K Beifuss, Tina L Gumienny

Abstract

C. elegans has proven to be a valuable model system for the discovery and functional characterization of many genes and gene pathways. More sophisticated tools and resources for studies in this system are facilitating continued discovery of genes with more subtle phenotypes or roles. Here we present a generalized protocol we adapted for identifying C. elegans genes with postembryonic phenotypes of interest using RNAi. This procedure is easily modified to assay the phenotype of choice, whether by light or fluorescence optics on a dissecting or compound microscope. This screening protocol capitalizes on the physical assets of the organism and molecular tools the C. elegans research community has produced. As an example, we demonstrate the use of an integrated transgene that expresses a fluorescent product in an RNAi screen to identify genes required for the normal localization of this product in late stage larvae and adults. First, we used a commercially available genomic RNAi library with full-length cDNA inserts. This library facilitates the rapid identification of multiple candidates by RNAi reduction of the candidate gene product. Second, we generated an integrated transgene that expresses our fluorecently tagged protein of i...Continue Reading

References

Feb 15, 1994·Proceedings of the National Academy of Sciences of the United States of America·M D YandellW B Wood
Nov 6, 1998·Nature·L Timmons, A Fire
Aug 15, 2002·Current Biology : CB·Femke SimmerRonald H A Plasterk
Apr 30, 2003·Genesis : the Journal of Genetics and Development·Cathy Savage-DunnRichard W Padgett
Sep 22, 2006·Methods in Molecular Biology·Todd Lamitina
Dec 1, 2007·WormBook : the Online Review of C. Elegans Biology·Theresa Stiernagle
Apr 19, 2008·Cold Spring Harbor Symposia on Quantitative Biology·A V SamuelsonG Ruvkun
Jun 4, 2008·Nature Reviews. Genetics·Michael Boutros, Julie Ahringer
Aug 5, 2008·Nature Methods·Sumeet SarinOliver Hobert
Oct 28, 2008·Nature Genetics·Christian Frøkjaer-JensenErik M Jorgensen
Dec 11, 2008·Journal of Visualized Experiments : JoVE·Laura A BerkowitzKim A Caldwell
Apr 29, 2010·British Journal of Pharmacology·Jean Giacomotto, Laurent Ségalat
Mar 10, 2011·Genetics·Jimmy J Zhuang, Craig P Hunter
Mar 25, 2011·Cellular and Molecular Life Sciences : CMLS·Rosina Giordano-Santini, Denis Dupuy

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Citations

Feb 9, 2016·Cell Reports·Ksenia Smurova, Benjamin Podbilewicz

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