RNAi-mediated inhibition of gene function in the follicle cell layer of the Drosophila ovary

Abstract

RNA-mediated interference (RNAi) has been reported to be an effective reverse genetic approach for studying gene function in various organisms. To assess RNAi as a means of examining genes expressed in ovarian follicle cells for their involvement in embryonic dorsal-ventral patterning, we tested the ability of transgenically expressed double-stranded RNA (dsRNA) directed against the dorsal group gene windbeutel to generate phenotypic effects in the progeny of expressing females. We observed that expression in follicle cells under the control of Gal4 transcribed from the strong and widely expressed alphaTub84B or Actin5C promoters led to efficient dorsalization of progeny embryos. Surprisingly, a variety of strongly expressed follicle cell-specific Gal4 enhancer trap lines failed to elicit an RNAi phenotype in combination with the windbeutel-specific dsRNA. These results stress the importance of careful choice of expression system and of conditions for use in transgenic RNAi-mediated studies of gene function. © 2004 Wiley-Liss, Inc.