Huang Jia-Hsin, Liu Yun, Lin Yu-Hsien, Belles Xavier, Lee How-Jing
Department of Entomology, National Taiwan University.
Department of Entomology, National Taiwan University; Biology Centre, Institute of Entomology, Czech Academy of Sciences; Faculty of Science, University of South Bohemia.
J Vis Exp. 2018 May 1(135):57385. doi: 10.3791/57385.
RNA interference (RNAi) has been widely applied for uncovering the biological functions of numerous genes, and has been envisaged as a pest control tool operating by disruption of essential gene expression. Although different methods, such as injection, feeding, and soaking, have been reported for successful delivery of double-stranded RNA (dsRNA), the efficiency of RNAi through oral delivery of dsRNA is highly variable among different insect groups. The German cockroach, Blattella germanica, is highly sensitive to the injection of dsRNA, as shown by many studies published previously. The present study describes a method to demonstrate that the dsRNA encapsulated with liposome carriers is sufficient to retard the degradation of dsRNA by midgut juice. Notably, the continuous feeding of dsRNA encapsulated by liposomes significantly reduces the tubulin expression in the midgut, and led to the death of cockroaches. In conclusion, the formulation and utilization of dsRNA lipoplexes, which protect dsRNA against nucleases, could be a practical use of RNAi for insect pest control in the future.
RNA干扰(RNAi)已被广泛应用于揭示众多基因的生物学功能,并被设想为一种通过破坏必需基因表达来控制害虫的工具。尽管已报道了不同的方法,如注射、喂食和浸泡,用于成功递送双链RNA(dsRNA),但通过口服递送dsRNA的RNAi效率在不同昆虫群体中差异很大。德国小蠊(Blattella germanica)对注射dsRNA高度敏感,此前发表的许多研究都表明了这一点。本研究描述了一种方法,以证明用脂质体载体包裹的dsRNA足以延缓中肠液对dsRNA的降解。值得注意的是,持续喂食脂质体包裹的dsRNA可显著降低中肠微管蛋白的表达,并导致蟑螂死亡。总之,保护dsRNA免受核酸酶作用的dsRNA脂质复合物的制备和应用,可能成为未来RNAi在害虫防治中的一种实际应用。
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