MicroRNA-based antiviral transgenic mosquitoes Aedes aegypti to suppress dengue virus replication
Date Issued
2016
Date
2016
Author(s)
Hsu, Chia-Wei
Abstract
Dengue is an arthropod-borne viral disease prevalent in tropical and subtropical regions, caused by the four dengue virus serotypes (DENV 1–4), which are transmitted by Aedes mosquitoes. Until now, effective vaccine and positive treatment are not available, and the use of insecticides also leads to resistance in mosquito vectors. Therefore, the alternative strategies for mosquito vector control are needed. Genetic manipulation via RNA interference (RNAi) in the mosquito can permanently reduce vector competence and subsequent transmission of DENV to the human. Recent studies have found that expression of virus-specific double stranded RNAs is a potential way to block the DENV replication and vector competence to virus. In this study, using the mariner (Mos1) transposase and genomic integration system, we generated serial transgenic mosquito lines which express anti-viral microRNAs under the control of A. aegypti poly-ubiquitin (Pub), serine-type enodpeptidase (AAEL001703) and carboxypeptidase A (CPA) promoters. We verified the integration of the transgene in the mosquitoes by qPCR and Southern blot. By using virus challenge with viremic bloodmeal, the viral titer and infection rate was significantly suppressed in the transgenic mosquitoes of CPA>8-miRNA, Pub>8-miRNA and AAEL001703>4-miRNA at day-7 post bloodmeal, compared to those of wild type mosquitoes. Repeat bloodmeal on CPA>8-miRNA provided effective antiviral efficiency at day-14 post infection. Transmission in vitro of virus from CPA>8-miRNA line was significantly diminished when compared to Pub>8-miRNA and control mosquitoes at day-14 post bloodmeal. Our study provides an evidence for DENV control strategy which suppress viral replication via genetic manipulation of miRNA in Aedes aegypti.
Subjects
DENV
Aedes aegypti
anti-viral miRNA
transgenic mosquitoes
virus challenge
repeat bloodmeal
Type
thesis
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