Breaking Barriers to Mosquito RNAi Control: A dsRNA Biomanufacturing and Nanodelivery Platform for Sustainable Vector Management

Abstract RNA interference (RNAi) offers a promising approach for pest control but is often limited by the instability of double-stranded RNA (dsRNA) and inefficient cellular delivery. We developed a biodegradable, redox-responsive polyaminoglycoside nanocarrier (SS-HPT) to improve dsRNA stability, cellular uptake, and intracellular release. SS-HPT readily formed stable nanocomplexes (∼300 nm, positively charged) that resisted nuclease degradation under simulated gut conditions and remained stable during storage. Fluorescence imaging demonstrated efficient uptake and widespread distribution in mosquito larvae. Intracellular disulfide cleavage promoted dsRNA release, enhancing gene silencing. Delivery of β-tubulin-targeting dsRNA via SS-HPT resulted in over 95% larval mortality at substantially lower doses than unprotected dsRNA. These findings establish SS-HPT as an effective platform for RNAi-based pest management.