Exon-specific RNAi: a tool for dissecting the functional relevance of alternative splicing.
The goal of functional genomics is to determine the function of each protein encoded by an organism. Typically, this is done by inactivating individual genes and, subsequently, analyzing the phenotype of the modified organisms. In higher eukaryotes, where a tremendous amount of alternative splicing occurs, such approaches are not feasible because they have the potential to simultaneously affect multiple proteins that could have quite distinct and important functions. Thus, it is necessary to develop techniques that inactivate only a subset of proteins synthesized from genes encoding alternatively spliced mRNAs. Here we demonstrate that RNA interference (RNAi) can be used to selectively degrade specific alternatively spliced mRNA isoforms in cultured Drosophila cells. This is achieved by treating the cells with double-stranded RNA corresponding to an alternatively spliced exon. This technique may prove to be a powerful tool to assess the function of proteins synthesized from alternatively spliced mRNAs. In addition, these results have implications regarding the mechanism of RNAi in Drosophila.
Celotto
Alicia M
AM
University of Connecticut Health Center, Department of Genetics and Developmental Biology, Farmington 06030-3301, USA.
Graveley
Brenton R
BR
D017398:Alternative Splicing; D000818:Animals; D004330:Drosophila; D005091:Exons; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger