Novel genomic cDNA hybrids produce effective RNA interference in adult Drosophila.
Drosophila melanogaster has been a premier genetic model system for nearly 100 years, yet lacks a simple method to disrupt gene expression. Here, we show genomic cDNA fusions predicted to form double-stranded RNA (dsRNA) following splicing, effectively silencing expression of target genes in adult transgenic animals. We targeted three Drosophila genes: lush, white, and dGq(alpha). In each case, target gene expression is dramatically reduced, and the white RNAi phenotype is indistinguishable from a deletion mutant. This technique efficiently targets genes expressed in neurons, a tissue refractory to RNAi in C. elegans. These results demonstrate a simple strategy to knock out gene function in specific cells in living adult Drosophila that can be applied to define the biological function of hundreds of orphan genes and open reading frames.
Kalidas
Savitha
S
Department of Pharmacology and Center for Basic Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
Smith
Dean P
DP
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D019677:Artificial Gene Fusion; D018076:DNA, Complementary / Q000235:genetics*; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D018390:Gene Targeting; D016678:Genome; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics / Q000502:physiology; D019476:Insect Proteins / Q000235:genetics; D009474:Neurons / Q000502:physiology; D009833:Olfactory Pathways / Q000502:physiology; D012313:RNA / Q000502:physiology*; D012330:RNA, Double-Stranded / Q000502:physiology; D018035:Receptors, Odorant / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology
D018528:ATP-Binding Cassette Transporters; D018076:DNA, Complementary; D029721:Drosophila Proteins; D005136:Eye Proteins; C097161:G protein a49B, Drosophila; D019476:Insect Proteins; D012330:RNA, Double-Stranded; D018035:Receptors, Odorant; C045648:odorant-binding protein; C059254:w protein, Drosophila; D012313:RNA; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins