Silencing of Toll pathway components by direct injection of double-stranded RNA into Drosophila adult flies.
Double-stranded RNA (dsRNA) gene interference is an efficient method to silence gene expression in a sequence-specific manner. Here we show that the direct injection of dsRNA can be used in adult Drosophila flies to disrupt function of endogenous genes in vivo. As a proof of principle, we have used this method to silence components of a major signaling cascade, the Toll pathway, which controls fruit fly resistance to fungal and Gram-positive bacterial infections. We demonstrate that the knockout is efficient only if dsRNA is injected in 4- or more day-old flies and that it lasts for at least 1 week. Furthermore, we report dsRNA-based epistatic gene analysis via injection of a mixture of two dsRNAs and propose that injection of dsRNA represents a powerful method for rapid functional analysis of genes in Drosophila melanogaster adults, particularly of those whose mutations are lethal during development.
Goto
Akira
A
UPR 9022 du Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, F-67084 Strasbourg Cedex, France.
Blandin
Stéphanie
S
Royet
Julien
J
Reichhart
Jean-Marc
JM
Levashina
Elena A
EA
D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D004843:Epistasis, Genetic; D005260:Female; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D010641:Phenotype; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000008:administration & dosage / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D015843:Serpins / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D013997:Time Factors; D051193:Toll-Like Receptors; D014157:Transcription Factors / Q000235:genetics
D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; C085443:Opa protein, Drosophila; D012330:RNA, Double-Stranded; D011956:Receptors, Cell Surface; D015843:Serpins; C068721:Tl protein, Drosophila; D051193:Toll-Like Receptors; D014157:Transcription Factors; C417282:nec protein, Drosophila; D049452:Green Fluorescent Proteins