RNA interference from a CAX trinucleotide repeat

• Published in: Genesis (New York, N.Y. : 2000) Vol. 34; no. 1-2; pp. 156 - 159
• Main Authors: Akten, Bikem, Suh, Joowon, Genova, Ginka K., Roberts, Mary A., Jackson, F. Rob
• Format: Journal Article
• Language: English
• Published: New York Wiley Subscription Services, Inc., A Wiley Company 01-09-2002
• Subjects: Animals; Animals, Genetically Modified; Crosses, Genetic; DNA-Binding Proteins; Drosophila melanogaster - genetics; Drosophila Proteins - genetics; Enhancer Elements, Genetic; Female; Gene Duplication; Membrane Proteins - genetics; RNA Interference; Saccharomyces cerevisiae Proteins - genetics; Sequence Analysis, DNA; Transcription Factors - genetics; Trinucleotide Repeats; Wings, Animal - abnormalities
• ISSN: 1526-954X; 1526-968X
• DOI: 10.1002/gene.10126

The use of RNA interference (RNAi) techniques, combined with a spatially controlled expression of double-stranded RNA (dsRNA), provides a convenient method for producing cell-type-specific perturbations of gene function. While studying the function of the Drosophila miniature-dusky (m-dy) gene complex, we produced transgenic flies expressing a dsRNA that would permit silencing of the dy gene (P{UAS-dy-dsRNA}). The m and dy genes share sequence similarity and both encode fly Cuticulin proteins that are required at the pupal stage of development for normal wing differentiation. In the absence of m-dy products, the wing is reduced in size, due to an autonomous effect on the size and/or shape of wing epidermal cells. Although a reduction in wing size is the only obvious morphological phenotype in m-dy flies, certain mutants exhibit reduced viability and/or fertility. To examine the phenotypic effects of the UAS-dy-dsRNA transgene, we ectopically expressed it within wing tissues, taking advantage of four different existing Drosophila GAL4 driver strains: P{GawB}30A, P{GawB}34B, MS1096, and A9. The first two strains are described in Flybase, the last two were obtained from M. O'Connor (Univ. of Minnesota). All four strains provide expression in wing and other imaginal discs.