Effectiveness of specific RNA-mediated interference through ingested double-stranded RNA in Caenorhabditis elegans.docVIP

/2000/2/1/research/0002.1 Research Effectiveness of specific RNA-mediated interference through ingested double-stranded RNA in Caenorhabditis elegans Ravi S Kamath, Maruxa Martinez-Campos, Peder Zipperlen, Andrew G Fraser and Julie Ahringer Address: Wellcome/CRC Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK. Correspondence: Julie Ahringer. E-mail: jaa@mole.bio.cam.ac.uk Published: 20 December 2000 Received: 12 September 2000 Revised: 10 October 2000 Accepted: 10 November 2000 Genome Biology 2000, 2(1):research0002.1–0002.10 The electronic version of this article is the complete one and can be found online at /2000/2/1/research/0002 ? GenomeB (Print ISSN 1465-6906; Online ISSN 1465-6914) Abstract Background: In Caenorhabditis elegans, injection of double-stranded RNA (dsRNA) results in the specific inactivation of genes containing homologous sequences, a technique termed RNA- mediated interference (RNAi). It has previously been shown that RNAi can also be achieved by feeding worms Escherichia coli expressing dsRNA corresponding to a specific gene; this mode of dsRNA introduction is conventionally considered to be less efficient than direct injection, however, and has therefore seen limited use, even though it is considerably less labor-intensive. Results: Here we present an optimized feeding method that results in phenotypes at least as strong as those produced by direct injection of dsRNA for embryonic lethal genes, and stronger for genes with post-embryonic phenotypes. In addition, the interference effect generated by feeding can be titrated to uncover a series of hypomorphic phenotypes informative about the functions of a given gene. Using this method, we screened 86 random genes on consecutive cosmids and identified functions for 13 new genes. These included two genes producing an uncoordinated phenotype (a previously