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dc.contributor.authorKondo, Shu
dc.contributor.authorBooker, Matthew
dc.contributor.authorPerrimon, Norbert
dc.date.accessioned2019-09-21T09:03:07Z
dc.date.issued2009
dc.identifier.citationKondo, Shu, Matthew Booker, and Norbert Perrimon. 2009. “Cross-Species RNAi Rescue Platform inDrosophila Melanogaster.” Genetics 183 (3): 1165–73. https://doi.org/10.1534/genetics.109.106567.
dc.identifier.issn0016-6731
dc.identifier.issn1943-2631
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:41384479*
dc.description.abstractRNAi-mediated gene knockdown in Drosophila melanogaster is a powerful method to analyze loss-of-function phenotypes both in cell culture and in vivo. However, it has also become clear that false positives caused by off-target effects are prevalent, requiring careful validation of RNAi-induced phenotypes. The most rigorous proof that an RNAi-induced phenotype is due to loss of its intended target is to rescue the phenotype by a transgene impervious to RNAi. For large-scale validations in the mouse and Caenorhabditis elegans, this has been accomplished by using bacterial artificial chromosomes (BACs) of related species. However, in Drosophila, this approach is not feasible because transformation of large BACs is inefficient. We have therefore developed a general RNAi rescue approach for Drosophila that employs Cre/loxP-mediated recombination to rapidly retrofit existing fosmid clones into rescue constructs. Retrofitted fosmid clones carry a selection marker and a phiC31 attB site, which facilitates the production of transgenic animals. Here, we describe our approach and demonstrate proof-of-principle experiments showing that D. pseudoobscura fosmids can successfully rescue RNAi-induced phenotypes in D. melanogaster both in cell culture and in vivo. Altogether, the tools and method that we have developed provide a gold standard for validation of Drosophila RNAi experiments.
dc.language.isoen_US
dc.publisherGenetics Society of America
dash.licenseMETA_ONLY
dc.titleCross-Species RNAi Rescue Platform in Drosophila melanogaster
dc.typeJournal Article
dc.description.versionVersion of Record
dc.relation.journalGenetics
dash.depositing.authorPerrimon, Norbert::4e34fce71a186b488b1846b199007bfb::600
dc.date.available2019-09-21T09:03:07Z
dash.workflow.comments1Science Serial ID 41965
dc.identifier.doi10.1534/genetics.109.106567
dash.source.volume183;3
dash.source.page1165


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