New Insights into Expanding the Insecticidal Spectrum of dsRNA Mediated by the High Sequence Identity between dsRNA and Nontarget mRNA

RNA interference (RNAi) is being used to develop methods to control pests, yet its widespread application is limited by the high comprehensive application cost of dsRNAs. Here, we utilized the high identity matching between double-stranded RNA (dsRNA) and nontarget genes to achieve expanding the dsRNA insecticidal spectrum. First, we found that dsRNA was more likely to induce off-target effects in genes with higher transcript levels and higher sequence identity; the existence of either a completely contiguous matching sequence exceeding 15 nt or a partially contiguous matching sequence of 24 nt between genes can lead to off-target effects in <i>Tribolium castaneum</i>. Accordingly, we successfully interfered with <i>T. castaneum</i> and <i>Laodelphax striatellus</i> using dsRNA targeted against <i>Nilaparvata lugens</i>. Additionally, the use of dsRNA targeting <i>L. striatellus</i> effectively interfered with <i>N. lugens</i>, both instances resulting in lethal effects. Moreover, the dsRNA spray method proved to be more efficient than the rice seedling soaking method to deliver dsRNA. Our research offers new insights into expanding the insecticidal spectrum of dsRNA mediated by a high degree of sequence identity between genes.