Location: Mosquito and Fly ResearchTitle: Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA) Author
Submitted to: Journal of Applied Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/29/2012
Publication Date: 7/1/2012
Citation: Coy, M.R., Sanscrainte, N.D., Chalaire, K.C., Inberg, A., Maayan, I., Glick, E., Paldi, N., Becnel, J.J. 2012. Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA. Journal of Applied Entomology. 136:741-748. Interpretive Summary: In order to determine if orally delivered double-stranded RNA (dsRNA) would reduce gene expression in adult Aedes aegypti mosquitoes as it has been shown in other insects, a method to deliver dsRNA to the mosquitoes in a sucrose solution supplied in a cotton wick was devised by scientists at the Center for Medical, Agricultural, and Veterinary Entomology, in Gainesville, FL and at Beelogics, LLC. A gut-expressed gene, vacuolar ATPase was the experimental target, and it was determined that gene expression could be reduced by delivery of dsRNA in this manner. This methodology will be used to test future targets to identify genes that can be combined with other control efforts to reduce mosquito populations.
Technical Abstract: The induction of the naturally occurring phenomenon of RNA interference (RNAi) to study gene function in insects is now common practice. With appropriately chosen targets, the RNAi pathway has also been exploited for insect control, typically through oral delivery of dsRNA. To determine if such an approach could be used in the yellow fever mosquito, Aedes aegypti, we adapted existing methods for the oral delivery of foreign compounds, such as amino acids and pesticides, to the mosquito through a sucrose solution. Using a non-specific dsRNA construct, we found that adult Ae. aegypti ingested dsRNA through this method, and that the ingested dsRNA can be recovered from the mosquitoes post-feeding. Through the feeding of a species-specific dsRNA construct against vacuolar ATPase, subunit A, we found that significant gene knockdown could be achieved at 12, 24 and 48 hours post-feeding.