Self-Assembling Nanoparticle Enhances RNA Interference Efficiency against Hyphantria Cunea (Drury) with Minimal Risk to Nontarget Organisms.

RNA interference (RNAi) represents a promising approach for insect pest management; however, its application in Lepidoptera is constrained by double-stranded RNA (dsRNA) instability, limited cellular uptake, and inefficient RNAi machinery. In this study, we developed a bacteriophage MS2 virus-like particle (VLP)-based delivery platform for hairpin RNA (hpRNA) targeting the invasive pest Hyphantria cunea. When expressed in Escherichia coli, MS2 VLPs efficiently encapsulate hpRNA, markedly enhancing its resistance to nuclease activity and environmental degradation. In addition, surface display of the HIV trans-activator of transcription (TAT) peptide on MS2 VLPs significantly improved cellular internalization of hpRNA, resulting in robust RNAi-mediated gene silencing in H. cunea at low hpRNA doses. Importantly, no adverse effects were detected in three nontarget organisms: Clostera restitura, Plagiodera versicolora, and the parasitoid Chouioia cunea. Together, these results demonstrate that the MS2-hpRNA system represents a scalable, effective, and environmentally safe strategy for RNA-based pest control.