Drosophila RNA Interference (RNAi) Using a Gal-4 Inducible Transgene Vector.

ABSTRACT

INTRODUCTIONRNA interference (RNAi) is a powerful method for determining the role of specific genes during Drosophila embryogenesis. This protocol describes a method for RNAi in vivo using tissue-specific Gal-4 transgenes to induce dsRNA synthesis from an upstream activator sequence (UAS) vector. This vector contains the desired exonic inverted sequences representing the target gene (preferably more than 400 bp) separated by a unique spacer, the first intron of the actin 5C gene. The inverted repeats are stable during cloning in E. coli with this intronic spacer and the intron is spliced out to produce an almost perfect dsRNA target for Dicer cleavage and the production of siRNAs.