Carroll Elijah, Kunte Nitish, McGraw Erin, Gautam Sujan, Range Ryan, Noveron-Nunez Jose A, Held David W, Avila L Adriana
Department of Entomology and Plant Pathology, Auburn University, Auburn, AL, United States.
Department of Biological Sciences, Auburn University, Auburn, AL, United States.
Front Insect Sci. 2023 May 12;3:1151789. doi: 10.3389/finsc.2023.1151789. eCollection 2023.
Gene silencing by feeding double-stranded (dsRNA) holds promise as a novel pest management strategy. Nonetheless, degradation of dsRNA in the environment and within the insect gut, as well as inefficient systemic delivery are major limitations to applying this strategy. Branched amphiphilic peptide capsules (BAPCs) complexed with dsRNA have been used to successfully target genes outside and inside the gut epithelium upon ingestion. This suggests that BAPCs can protect dsRNA from degradation in the gut environment and successfully shuttle it across gut epithelium. In this study, our objectives were to 1) Determine whether feeding on BAPC-dsRNA complexes targeting a putative peritrophin gene of would result in the suppression of gut peritrophin synthesis, and 2) gain insight into the cellular uptake mechanisms and transport of BAPC-dsRNA complexes across the larval midgut of Our results suggest that BAPC-dsRNA complexes are readily taken up by the midgut epithelium, and treatment of the tissue with endocytosis inhibitors effectively suppresses intracellular transport. Further, assessment of gene expression in BAPC- peritrophin dsRNA fed beetles demonstrated significant downregulation in mRNA levels relative to control and/or dsRNA alone. Our results demonstrated that BAPCs increase the efficacy of gene knockdown relative to dsRNA alone in adults. To our knowledge, this is the first report on nanoparticle-mediated dsRNA delivery through feeding in .
通过喂食双链RNA（dsRNA）实现基因沉默有望成为一种新型害虫管理策略。尽管如此，dsRNA在环境和昆虫肠道内的降解以及低效的全身递送是应用该策略的主要限制因素。与dsRNA复合的支链两亲肽胶囊（BAPCs）在摄入后已成功靶向肠道上皮细胞内外的基因。这表明BAPCs可以保护dsRNA在肠道环境中不被降解，并成功地将其穿梭穿过肠道上皮细胞。在本研究中，我们的目标是：1）确定喂食靶向假定的围食膜蛋白基因的BAPC-dsRNA复合物是否会导致肠道围食膜蛋白合成受到抑制，以及2）深入了解BAPC-dsRNA复合物在幼虫中肠的细胞摄取机制和转运。我们的结果表明，中肠上皮细胞很容易摄取BAPC-dsRNA复合物，并且用内吞作用抑制剂处理组织可有效抑制细胞内转运。此外，对喂食BAPC-围食膜蛋白dsRNA的甲虫的基因表达评估表明，相对于对照和/或单独的dsRNA，mRNA水平显著下调。我们的结果表明，相对于单独的dsRNA，BAPCs提高了在成虫中基因敲低的效果。据我们所知，这是关于通过喂食在中进行纳米颗粒介导的dsRNA递送的首次报道。
