Targeted disruption of gene function in Drosophila by RNA interference (RNA-i): a role for nautilus in embryonic somatic muscle formation.
The expression of the MyoD gene homolog, nautilus (nau), in the Drosophila embryo defines a subset of mesodermal cells known as the muscle "pioneer" or "founder" cells. These cells are thought to establish the future muscle pattern in each hemisegment. Founders appear to recruit fusion-competent mesodermal cells to establish a particular muscle fiber type. In support of this concept every somatic muscle in the embryo is associated with one or more nautilus-positive cells. However, because of the lack of known (isolated) nautilus mutations, no direct test of the founder cell hypothesis has been possible. We now have utilized toxin ablation and genetic interference by double-stranded RNA (RNA interference or RNA-i) to determine both the role of the nautilus-expressing cells and the nautilus gene, respectively, in embryonic muscle formation. In the absence of nautilus-expressing cells muscle formation is severely disrupted or absent. A similar phenotype is observed with the elimination of the nautilus gene product by genetic interference upon injection of nautilus double-stranded RNA. These results define a crucial role for nautilus in embryonic muscle formation. The application of RNA interference to a variety of known Drosophila mutations as controls gave phenotypes essentially indistinguishable from the original mutation. RNA-i provides a powerful approach for the targeted disruption of a given genetic function in Drosophila.
Misquitta
L
L
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Paterson
B M
BM
D000818:Animals; D002823:Chorion; D004330:Drosophila / Q000196:embryology* / Q000235:genetics*; D029721:Drosophila Proteins; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000502:physiology; D004627:Embryonic Induction; D005260:Female; D018507:Gene Expression Regulation, Developmental; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva; D009124:Muscle Proteins; D009132:Muscles / Q000196:embryology*; D016296:Mutagenesis; D010641:Phenotype; D016372:RNA, Antisense / Q000494:pharmacology*; D012330:RNA, Double-Stranded / Q000235:genetics; D014157:Transcription Factors