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

摘要: Whole-genome transgenic RNAi libraries permit systematic genetic screens in individual tissues of Drosophila. However, there is a high incidence nonspecific phenotypes because off-target effects. To minimize such effects, it essential to obtain deeper understanding the specificity action RNAi. Here, vivo assays are used determine minimum, contiguous nucleotide pairing required between an siRNA and target mRNA generate phenotype. We observe that as few 16 nucleotides homology sufficient attenuate gene activity. This finding provides explanation for effects observed RNAi-based screens. Toward improving efficacy RNAi-induced vivo, we describe expression vectors allow coexpression one or more siRNAs with fluorescent reporter cultured cells flies. system makes use small intron from ftz segmentation provide efficient processing synthetic transcript. These studies foundation specific effective silencing