Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti
作者： A. W. E. Franz , I. Sanchez-Vargas , Z. N. Adelman , C. D. Blair , B. J. Beaty
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摘要: Mosquitoes (Aedes aegypti) were genetically modified to exhibit impaired vector competence for dengue type 2 viruses (DENV-2). We exploited the natural antiviral RNA interference (RNAi) pathway in mosquito midgut by constructing an effector gene that expresses inverted-repeat (IR) derived from premembrane protein coding region of DENV-2 genome. The A. aegypti carboxypeptidase A promoter was used express IR epithelial cells after ingestion a bloodmeal. and inserted into genome white-eye Puerto Rico Rexville D (Higgs’ white eye) strain using nonautonomous mariner MosI transformation system. transgenic family, Carb77, expressed Carb77 mosquitoes ingesting artificial bloodmeal containing exhibited marked reduction viral envelope antigen midguts salivary glands infection. titration individual showed most poorly supported virus replication. Transmission vitro line significantly diminished when compared control mosquitoes. presence DENV-2-derived siRNAs extracts loss resistance phenotype RNAi interrupted proved caused response. Engineering show high level against provides powerful tool developing population replacement strategies transmission viruses.
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