Description:
<jats:p>RNA interference (RNAi) is a promising next generation technology for the development of species-specific pest management. The key to successful RNAi based-plant protection is dependent in part on data-driven target gene selection, a challenging task due to the absence of laboratory strains and the seasonality of most pest species. In this study, we aimed to identify novel target genes by performing a knowledge-based approach in order to expand the spectrum of known potent RNAi targets. Recently, the protein-coding genes <jats:italic>ncm, Rop, RPII-140</jats:italic>, and <jats:italic>dre4</jats:italic> have been identified as sensitive RNAi targets for pest control. Based on these potent RNAi targets, we constructed an interaction network and analyzed a selection of 30 genes in the model beetle <jats:italic>Tribolium castaneum</jats:italic> via injection of dsRNA synthesized by <jats:italic>in vitro</jats:italic> transcription. Nineteen of these targets induced significant mortality of over 70%, including six that caused 100% lethality. Orthologs of active <jats:italic>T. castaneum</jats:italic> RNAi targets were verified in the economically important coleopteran pests <jats:italic>Diabrotica virgifera virgifera</jats:italic> and <jats:italic>Brassicogethes aeneus</jats:italic>. Knockdown of <jats:italic>D. v. virgifera</jats:italic> genes coding for transcription factor <jats:italic>Spt5, Spt6</jats:italic>, and RNA polymerase II subunit RPII-33 caused over 90% mortality in larval feeding assays. Injection of dsRNA constructs targeting RPII-215 or the pre-mRNA-processing factor Prp19 into adult <jats:italic>B. aeneus</jats:italic> resulted in high lethality rates of 93 and 87%, respectively. In summary, the demonstrated knowledge-based approaches increased the probability of identifying novel lethal RNAi target genes from 2% (whole genome) to 36% (transcription- and splicing-related genes). In addition, performing RNAi pre-screening in a model insect increased also the probability of the identification essential genes in the difficult-to-work-with pest species <jats:italic>D. v. virgifera</jats:italic> and <jats:italic>B. aeneus</jats:italic>.</jats:p>