Vectors and parameters that enhance the efficacy of RNAi-mediated gene disruption in transgenic Drosophila.

ABSTRACT: Whole-genome transgenic RNAi libraries permit systematic genetic screens in individual tissues of Drosophila. However, there is a high incidence of nonspecific phenotypes because of off-target effects. To minimize such effects, it is essential to obtain a deeper understanding of the specificity of action of RNAi. Here, in vivo assays are used to determine the minimum, contiguous nucleotide pairing required between an siRNA and a target mRNA to generate a phenotype. We observe that as few as 16 nucleotides of contiguous homology are sufficient to attenuate gene activity. This finding provides an explanation for the high incidence of off-target effects observed in RNAi-based genetic screens. Toward improving the efficacy of RNAi-induced phenotypes in vivo, we describe siRNA expression vector

SUBMITTER: Haley B 

PROVIDER: S-EPMC2895090 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature