Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans.
Genetic interference mediated by double-stranded RNA (RNAi) has been a valuable tool in the analysis of gene function in Caenorhabditis elegans. Here we report an efficient induction of RNAi using bacteria to deliver double-stranded RNA. This method makes use of bacteria that are deficient in RNaseIII, an enzyme that normally degrades a majority of dsRNAs in the bacterial cell. Bacteria deficient for RNaseIII were engineered to produce high quantities of specific dsRNA segments. When fed to C. elegans, such engineered bacteria were found to produce populations of RNAi-affected animals with phenotypes that were comparable in expressivity to the corresponding loss-of-function mutants. We found the method to be most effective in inducing RNAi for non-neuronal tissue of late larval and adult hermaphrodites, with decreased effectiveness in the nervous system, in early larval stages, and in males. Bacteria-induced RNAi phenotypes could be maintained over the course of several generations with continuous feeding, allowing for convenient assessments of the biological consequences of specific genetic interference and of continuous exposure to dsRNAs.
Department of Embryology, Carnegie Institution of Washington, 115 West University Parkway, Baltimore, MD 21210, USA.
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D000818:Animals; D030801:Animals, Genetically Modified; D001419:Bacteria / Q000235:genetics / Q000254:growth & development; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D005260:Female; D005453:Fluorescence; D015870:Gene Expression; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008297:Male; D010641:Phenotype; D012330:RNA, Double-Stranded / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D014162:Transfection / Q000379:methods
D008164:Luminescent Proteins; D012330:RNA, Double-Stranded; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins