In vitro studies provide insight into effects of Dicer-2 helicase mutations in Drosophila melanogaster

Mutations in the helicase domain have differential effects on processing hpRNAs to endo-siRNAs. (A) Size distribution of endo-siRNAs from whole flies derived from esi-1 and esi-2 hpRNA loci normalized to miRNA reads, with graphs to the right showing magnification of indicated regions. Six libraries were analyzed for dcr-2^null and GFP::Dcr-2^F225G, and five libraries for other genotypes. (B) Levels of esi-1 (top) and esi-2 (bottom) hpRNA precursors as quantified by RT-qPCR in testes (n = 4, each with three biological replicates), normalized to the housekeeping gene RP49. Median and interquartile range are represented. All strains compared to GFP::Dcr-2^WT, using one-way ANOVA with Tukey correction for multiple comparison. (***) P < 0.001; (ns) not significant. Horizontal lines show statistics for additional comparisons: (#) P < 0.05; (##) P < 0.01; (###) P < 0.001; (ns) not significant.