Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway.
We investigated the potential of double-stranded RNA to interfere with the function of genes in Drosophila. Injection of dsRNA into embryos resulted in potent and specific interference of several genes that were tested. In contrast, single-stranded RNA weakly interfered with gene activity. The method was used to determine the reception mechanism of the morphogen Wingless. Interference of the frizzled and Drosophila frizzled 2 genes together produced defects in embryonic patterning that mimic loss of wingless function. Interference of either gene alone had no effect on patterning. Epistasis analysis indicates that frizzled and Drosophila frizzled 2 act downstream of wingless and upstream of zeste-white3 in the Wingless pathway. Our results demonstrate that dsRNA interference can be used to analyze many aspects of gene function.
Kennerdell
J R
JR
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
Carthew
R W
RW
D000818:Animals; D001426:Bacterial Proteins; D019521:Body Patterning / Q000235:genetics; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics*; D004843:Epistasis, Genetic; D051157:Frizzled Receptors; D051717:Fushi Tarazu Transcription Factors; D038362:Glycogen Synthase Kinase 3; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D007150:Immunohistochemistry; D007267:Injections; D019476:Insect Proteins / Q000235:genetics / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology*; D016372:RNA, Antisense / Q000235:genetics; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled; D017981:Receptors, Neurotransmitter; D012097:Repressor Proteins / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism; D051155:Wnt1 Protein
D001426:Bacterial Proteins; D029721:Drosophila Proteins; C063459:En protein, Drosophila; D051157:Frizzled Receptors; D051717:Fushi Tarazu Transcription Factors; D018398:Homeodomain Proteins; D019476:Insect Proteins; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D016372:RNA, Antisense; D012330:RNA, Double-Stranded; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D017981:Receptors, Neurotransmitter; D012097:Repressor Proteins; D014157:Transcription Factors; D051155:Wnt1 Protein; C052837:eve protein, Drosophila; C497169:ftz protein, Drosophila; C058784:fz protein, Drosophila; C101703:fz2 protein, Drosophila; C075878:ttk protein, Drosophila; C074622:wg protein, Drosophila; D017346:Protein Serine-Threonine Kinases; C080335:Sgg protein, Drosophila; D038362:Glycogen Synthase Kinase 3