Use of RNA interference to dissect the roles of trans-acting factors in alternative pre-mRNA splicing.
RNA interference (RNAi) is becoming a popular method for analyzing gene function in a variety of biological processes. We have used RNAi in cultured Drosophila cells to identify trans-acting factors that regulate the alternative splicing of endogenously transcribed pre-mRNAs. We have generated a dsRNA library comprising approximately 70% of the Drosophila genes encoding RNA binding proteins and assessed the function of each protein in the regulation of alternative splicing. This approach not only identifies trans-acting factors regulating specific alternative splicing events, but also can provide insight into the alternative splicing regulatory networks of Drosophila. Here, we describe this RNAi approach to identify alternative splicing regulatory proteins in detail.
Park
Jung W
JW
Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3301, USA.
Graveley
Brenton R
BR
D017398:Alternative Splicing; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000378:metabolism; D034622:RNA Interference; D012322:RNA Precursors / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000008:administration & dosage; D016601:RNA-Binding Proteins / Q000502:physiology; D015534:Trans-Activators / Q000502:physiology*
D029721:Drosophila Proteins; D012322:RNA Precursors; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D015534:Trans-Activators