A simplified miRNA-based gene silencing method for Drosophila melanogaster.
MicroRNA-based RNA interference is commonly used to produce loss-of-function phenotypes in mammalian systems, but is used only sparingly in invertebrates such as Caenorhabditis elegans and Drosophila melanogaster. Here, we evaluate this method in transgenic strains of D. melanogaster and cultured S2 cells. High throughput-ready expression vectors were developed that permit rapid cloning of synthetic hairpin RNAs. As proof of concept, this method was used for the efficient silencing of dpp gene activity in the adult wing, and the analysis of the general RNA Polymerase II (Pol II) elongation factor, Nelf-E.
Haley
Benjamin
B
Department of Molecular and Cell Biology, Division of Genetics, Genomics, and Development, Center for Integrative Genomics, University of California, Berkeley, Berkeley, California 94720, USA. benhaley@berkeley.edu
Hendrix
David
D
Trang
Vinh
V
Levine
Michael
M
D000818:Animals; D002460:Cell Line; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics*; D034622:RNA Interference; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D014921:Wings, Animal / Q000378:metabolism
D029721:Drosophila Proteins; D035683:MicroRNAs; D016601:RNA-Binding Proteins; D014157:Transcription Factors; C064196:dpp protein, Drosophila; C118437:negative elongation factor