Development of a web-based platform for designing effective and eco-friendly double-stranded RNA-based insecticides

RNA interference (RNAi) has emerged as an environmentally friendly alternative to conventional insecticides for pest control. RNAi-based pest control relies on the delivery of double-stranded RNA (dsRNA) complementary to essential genes, causing lethal and sublethal effects in the target pest. There is an enormous interest in developing dsRNA-based insecticides against diverse pest species, but a comprehensive pipeline is lacking. Here, we introduce a web-based platform that supports the design of dsRNA for RNAi-based pest control: dsRIP (set to launch at dsrip.uni-goettingen.de by November 2024). dsRIP offers three integrated tools. (A) RNAi target gene selector identifies the orthologs of highly effective target genes established through a genome-wide screen in Tribolium castaneum and subsequent testing on other insect pests. The tool readily supports more than 40 pest species and users can easily add new species of interest. (B) Lethal efficiency optimizer finds the best target region within gene sequences to maximize lethal effects. This tool was established by conducting bioassays with multiple dsRNAs to identify the sequence features capable of improving lethal effects in tandem with small RNA sequencing to get insights into dsRNA processing. (C) Off-target minimizer predicts the transcripts in non-target organisms, such as bees, targeted by dsRNA. The essential gene orthologs in the non-target organisms were predicted to especially minimize regions carrying high detrimental effect risks. These three tools guide the design of dsRNAs optimized for high insecticidal effects against pests while ensuring minimal harm to non-target organisms to improve RNAi-based pest control strategies.