Guiding RNAi Design Through Characterization of Endogenous Small RNA Pathways

RNA interference (RNAi) is a common eukaryotic gene regulation process driven by small RNA effectors. Mechanisms that govern regulatory small noncoding RNA behavior have been extensively described in only a handful of organisms, which suggests that the most effective RNAi approach in many organisms, such as insect pests, remains to be determined. Taking advantage of advances in high-throughput sequencing, characterization of small RNA molecules can be achieved through bioinformatic approaches without the need for genetic experiments. This chapter describes pipelines for characterizing three main classes of small RNAs (microRNAs, small-interfering RNAs, and piwi-associated RNAs) using computationally determined small RNA biogenesis signatures. Obtaining information regarding the abundance of different small RNA classes through these pipelines will lead to a better-informed RNAi strategy, thereby identifying the most efficacious approach for RNAi.