In Vivo RNAi Rescue in Drosophila melanogaster with Genomic Transgenes from Drosophila pseudoobscura
作者： Christoph C. H. Langer , Radoslaw K. Ejsmont , Cornelia Schönbauer , Frank Schnorrer , Pavel Tomancak
DOI: 10.1371/JOURNAL.PONE.0008928
关键词:
摘要: Background Systematic, large-scale RNA interference (RNAi) approaches are very valuable to systematically investigate biological processes in cell culture or tissues of organisms such as Drosophila. A notorious pitfall all RNAi technologies potential false positives caused by unspecific knock-down genes other than the intended target gene. The ultimate proof for specificity is a rescue construct immune RNAi, typically originating from related species. Methodology/Principal Findings We show that primary sequence divergence areas targeted Drosophila melanogaster hairpins five non-melanogaster species sufficient identify orthologs 81% predicted be refractory. We use clones genomic fosmid library pseudoobscura demonstrate phenotypes muscles. Four out we tested harbour cross-species functionality gene assayed, and three four phenotype melanogaster. Conclusions/Significance The designed seamless transgenesis can readily used systematic manner.
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