Abstract
The Aedes aegypti densovirus (AeDNV) has potential as a delivery vector for foreign nucleic acids into mosquito cells. In this study, we investigated the ability of plasmids containing recombinant viral transducing genome to induce RNA interference (RNAi) effects in C6/C36 cells. We then evaluated the efficiency of a recombinant AeDNV vector to induce RNAi in Aedes albopictus larvae. We found that the expression of V-ATPase was inhibited by up to 90% at 96 h post-transfection in transfected C6/C36 cells. In addition, the bioinsecticidal activities of various RNAi-expressing AeDNV vectors used to infect Ae. albopictus larvae were also tested. We found that when Ae. albopictus larvae were infected with recombinant AeDNV, expression of V-ATPase was downregulated by nearly 70% compared to controls. Furthermore, the median survival time bioassays demonstrated that recombinant AeDNV caused more serious pathogenic effects than the wild type virus. This is the first report showing that recombinant virus plasmid and corresponding recombinant AeDNV can be used as an effective in vitro and in vivo RNAi delivery system, respectively.
Highlights
Mosquito-borne diseases are a major international public health problem that continue to pose a public health threat [1]
non-structural 1 (NS1) protein is preserved in its entirety in the recombinant genome as the NS1GFP fusion protein, because of the multiple activities that this densovirus replication initiator protein possesses
green fluorescent protein (GFP) provides a robust marker for the recombinant vectortransfected cells in vitro and in vivo
Summary
Mosquito-borne diseases are a major international public health problem that continue to pose a public health threat [1]. Chemical insecticides, which have traditionally been used in response to epidemics, are a major part of sustainable, integrated mosquito management for the prevention of mosquito-borne diseases Such strategies have proven to be relatively ineffective or undesirable as a result of to the development of resistance within mosquito populations and the negative environmental impacts [2,3]. In light of these problems, the search for new alternative approaches that could be applied to combat the spread of these diseases continues. Microinjection is highly technically demanding and time-consuming, and is not suitable for high-throughput genetic analyses or practical applications, including mosquito control
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