In Vivo RNAi Rescue in Drosophila melanogaster with Genomic Transgenes from Drosophila pseudoobscura
| Autor: | Schnorrer, Frank, Tomancak, Pavel, Schönbauer, Cornelia, Ejsmont, Radoslaw K., Langer, Christoph C. H. |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
RNA-Interferenz
Schwarzbäuchige Taufliege Drosophila melanogaster Genomforschung Sequenz-Abgleich Larven RNA interference Drosophila melanogaster invertebrate genomics genomic library screening sequence alignment genomic library construction Larvae RNA stem-loop structure info:eu-repo/classification/ddc/610 ddc:610 |
| Druh dokumentu: | Článek |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0008928 |
| Popis: | Background Systematic, large-scale RNA interference (RNAi) approaches are very valuable to systematically investigate biological processes in cell culture or in tissues of organisms such as Drosophila. A notorious pitfall of all RNAi technologies are potential false positives caused by unspecific knock-down of genes other than the intended target gene. The ultimate proof for RNAi specificity is a rescue by a construct immune to RNAi, typically originating from a related species. Methodology/Principal Findings We show that primary sequence divergence in areas targeted by Drosophila melanogaster RNAi hairpins in five non-melanogaster species is sufficient to identify orthologs for 81% of the genes that are predicted to be RNAi refractory. We use clones from a genomic fosmid library of Drosophila pseudoobscura to demonstrate the rescue of RNAi phenotypes in Drosophila melanogaster muscles. Four out of five fosmid clones we tested harbour cross-species functionality for the gene assayed, and three out of the four rescue a RNAi phenotype in Drosophila melanogaster. Conclusions/Significance The Drosophila pseudoobscura fosmid library is designed for seamless cross-species transgenesis and can be readily used to demonstrate specificity of RNAi phenotypes in a systematic manner. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
| Externí odkaz: |
|