Abstract
Development of novel and specific insect pest management methods is critical for overcoming pesticide resistance and off-target effects. Gene silencing through consumption of double stranded (dsRNA) by insects shows promise in this area. Association of dsRNA with nanoparticles confers protection against nucleases, and can also promote translocation of dsRNA across the midgut epithelial cell membranes, and overall enhance gene knockdown effects. However, many mysteries of how nanoparticles and dsRNA are internalized by cells and subsequently transported across the midgut epithelium remain to be unraveled. In this article, we investigate the role of endocytosis and transcytosis in the uptake and transport of dsRNA and nanoparticles through midgut epithelium cells. Spodoptera frugiperda (Sf9) cells and branched amphiphilic peptide nano-capsules (BAPCs) were used as an experimental model. Additionally, analyses of reactive oxygen and nitrogen species (ROS/RNS) were performed to demonstrated that cell viability was minimally impacted by the BAPCs-dsRNA complex. Results suggests that clathrin-mediated endocytosis and macropinocytois are largely responsible for cellular uptake, and once within the midgut, transcytosis is involved in shuttling BAPCs from the lumen to the hemolymph. BAPCs were not found to be toxic to Sf9 cells or generate damaging reactive species once internalized. This opens up further possibilities for BAPCs as a new insect pest management method.
Insight Into Cellular Uptake and Transcytosis of Peptide Nanoparticles in a Generalist Caterpillar.
