Development of an Efficient Recombinant Mosquito Densovirus-Mediated RNA Interference System and Its Preliminary Application in Mosquito Control

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.

Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae

“…Another promising technology to conduct functional validation in aphids is the use of a gene vector derived from an aphid virus. In this respect, the Myzus persicae densovirus [78,79], which possesses a small single-stranded DNA genome, represents a good candidate following similar work with a mosquito densovirus [80]. This functional virus-based technology was used to inhibit gene expression, but also to over-express a specific gene, which would represent a major breakthrough in aphid post-genomic research.…”

New Mode of Action for a Knottin Protein Bioinsecticide

“…When ingested by insects, PA1b could block V-ATPase activity, and this would prevent the intestinal absorption of nutrients (28,29). Moreover, it has been demonstrated in the literature that targeting the V-ATPase of insects is a promising approach in the fight against insect pests (33). PA1b is a peptidic inhibitor of V-ATPase, so it could be used in transgenic approaches or in virus-mediated expression of the toxin for plant protection or to control mosquitoes (33,34).…”

RNA Interference for Mosquito and Mosquito-Borne Disease Control

“…Additionally, using a short hairpin RNA (shRNA) Densovirus expression system to silence ATPase subunit A significantly reduced the lifespan of Ae. albopictus larvae [81]. Variation in the target subunit could explain the variation of these results because not all V-ATPase genes are considered essential.…”

“…aegypti densovirus (AaDNV) genome was expressed in a plasmid containing a Pol III promoter-driven expression cassettes containing short hairpin RNA (shRNA) targeting Ae. aegypti V-ATPase subunit A [81]. Expression of the plasmid in Ae.…”