Lu Yun-Heng, Liu Yun, Lin Yu-Chun, Su Yu-Jyuan, Henry David, Lacarriere Valerie, Haller Samantha, Lin Yu-Hsien, Chiang Pei-Chen, Hsu Cheng-I, Hsu Wei-Lun, Chan Yi-Chieh, Chen Li, Wu Yueh-Lung
Department of Entomology, National Taiwan University, Taipei 106, Taiwan.
Corning European Technology Center, Avon, France.
Int J Biol Macromol. 2025 Aug;319(Pt 3):145091. doi: 10.1016/j.ijbiomac.2025.145091. Epub 2025 Jun 9.
RNA interference (RNAi) represents a promising biotechnological strategy for sustainable pest control by silencing essential insect genes, resulting in increased mortality. However, its practical application remains limited due to environmental instability and delivery inefficiency of RNA molecules. In this study, three novel RNA-delivery nanoparticles-Polyplex, CS-TPP, and Lipoplex- were developed and their ability to enhance RNA stability and gene silencing efficacy against the intestinal mesh gene in Spodoptera litura was evaluated. Among them, Lipoplex showed superior performance in protecting double-stranded RNA (dsRNA) and small interfering RNA (siRNA) from degradation caused by heat, UV radiation, and RNases. In vitro cellular uptake studies using the S. litura SL1A cell line revealed significantly enhanced siRNA internalization with Lipoplex compared to other formulations. In vivo bioassays demonstrated that siRNA-loaded Lipoplex efficiently suppressed mesh expression, impaired larval development, and caused high mortality rate in early-instar larvae. Furthermore, leaf bioassay confirmed Lipoplex's potential as a stable and effective delivery platform under environmental conditions. Our findings underscore the promise of Lipoplex nanoparticles as a robust nanocarrier system for RNAi-based pest control, offering both protection and efficient intracellular delivery of RNA molecules in target insect species.
RNA干扰（RNAi）是一种很有前景的生物技术策略，可通过沉默昆虫必需基因来实现可持续害虫防治，从而提高死亡率。然而，由于RNA分子的环境不稳定性和递送效率低下，其实际应用仍然有限。在本研究中，开发了三种新型RNA递送纳米颗粒——多聚体、壳聚糖-三聚磷酸钠和脂质体，并评估了它们增强RNA稳定性以及对斜纹夜蛾肠道网蛋白基因的基因沉默效果的能力。其中，脂质体在保护双链RNA（dsRNA）和小干扰RNA（siRNA）免受热、紫外线辐射和核糖核酸酶降解方面表现出卓越性能。使用斜纹夜蛾SL1A细胞系进行的体外细胞摄取研究表明，与其他制剂相比，脂质体显著增强了siRNA的内化。体内生物测定表明，负载siRNA的脂质体有效地抑制了网蛋白表达，损害了幼虫发育，并导致初龄幼虫的高死亡率。此外，叶片生物测定证实了脂质体在环境条件下作为稳定且有效递送平台的潜力。我们的研究结果强调了脂质体纳米颗粒作为基于RNAi的害虫防治强大纳米载体系统的前景，为目标昆虫物种中的RNA分子提供保护和高效的细胞内递送。
