There has been limited success in the usage of exogenous small interference RNA (siRNA) or small hairpin RNA (shRNA) to trigger RNA interference (RNAi) in insects. Instead, long double-stranded RNAs (dsRNA) are used to induce knockdown of target genes in insects. Here, we compared the potency of si/sh RNAs and dsRNA in Colorado potato beetle (CPB) cells. CPB cells showed highly efficient RNAi response to dsRNA. However, si/sh RNAs were inefficient in triggering RNAi in CPB cells. Confocal microscopy observations of Cy3 labeled-si/sh RNA cellular uptake revealed reduced si/sh RNA uptake compared to dsRNA. si/sh RNAs were stable in the conditioned media of CPB cells. Although in a small amount, when internalized by CPB cells, the si/sh RNAs were processed by the Dicer enzyme. Lipid-mediated transfection and chimeric dsRNA approaches were used to improve the delivery of si/sh RNAs. Our results suggest that the uptake of si/sh RNAs is inefficient in CPB cells, resulting in ineffective RNAi response. However, with the help of effective delivery methods, si/sh RNA could be a useful option for developing target-specific RNAi-mediated biopesticides.
|Journal||Archives of Insect Biochemistry and Physiology|
|State||Accepted/In press - 2023|
Bibliographical noteFunding Information:
Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award number R01GM070559 and the National Institute of Food and Agriculture, the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies (under grant no. IIP‐1821936, and by industry partners), and US Department of Agriculture (under HATCH Project 2351177000).
© 2023 Wiley Periodicals LLC.
- cellular uptake
ASJC Scopus subject areas
- Insect Science