- Titel
- In Vivo RNAi Rescue in Drosophila melanogaster with Genomic Transgenes from Drosophila pseudoobscura
- Zitierfähige Url:
- https://nbn-resolving.org/urn:nbn:de:bsz:14-qucosa-185372
- Quellenangabe
- PLoS ONE, Vol. 5 (2010) Bd. 1, e8928, ISSN 1932-6203
- Erstveröffentlichung
- 2010
- Abstract (EN)
- 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.
- Andere Ausgabe
- DOI: 10.1371/journal.pone.0008928
- Link zur Originalpublikation in der Zeitschrift PLoS ONE.
Link: http://dx.doi.org/10.1371/journal.pone.0008928 - Freie Schlagwörter (DE)
- RNA-Interferenz, Schwarzbäuchige Taufliege, Drosophila melanogaster, Genomforschung, Sequenz-Abgleich, Larven
- Freie Schlagwörter (EN)
- RNA interference, Drosophila melanogaster, invertebrate genomics, genomic library screening, sequence alignment, genomic library construction, Larvae, RNA stem-loop structure
- Klassifikation (DDC)
- 610
- Klassifikation (RVK)
- XA 10000
- Verlag
- PloS, Lawrence, Kan.
- URN Qucosa
- urn:nbn:de:bsz:14-qucosa-185372
- Veröffentlichungsdatum Qucosa
- 10.12.2015
- Dokumenttyp
- Artikel
- Sprache des Dokumentes
- Englisch
- Lizenz / Rechtehinweis
CC BY 4.0