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开发一种高效的重组蚊虫浓核病毒介导的 RNA 干扰系统及其在蚊虫控制中的初步应用。

Development of an efficient recombinant mosquito densovirus-mediated RNA interference system and its preliminary application in mosquito control.

机构信息

Key Laboratory of Prevention and Control of Emerging Infectious Diseases of Guangdong Higher Education Institutes, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, China.

出版信息

PLoS One. 2011;6(6):e21329. doi: 10.1371/journal.pone.0021329. Epub 2011 Jun 16.

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.

摘要

埃及伊蚊浓核病毒(AeDNV)有可能成为将外源核酸递送至蚊子细胞的载体。在这项研究中,我们研究了含有重组病毒转导基因组的质粒诱导 C6/C36 细胞中 RNA 干扰(RNAi)的能力。然后,我们评估了重组 AeDNV 载体在白纹伊蚊幼虫中诱导 RNAi 的效率。我们发现,转染的 C6/C36 细胞中转录后 96 小时,V-ATPase 的表达被抑制了多达 90%。此外,还测试了用于感染白纹伊蚊幼虫的各种表达 RNAi 的 AeDNV 载体的生物杀虫活性。我们发现,与对照组相比,重组 AeDNV 感染白纹伊蚊幼虫时,V-ATPase 的表达下调了近 70%。此外,中值存活时间生物测定表明,重组 AeDNV 比野生型病毒引起更严重的致病性影响。这是首次报道重组病毒质粒和相应的重组 AeDNV 分别可作为有效的体外和体内 RNAi 递送系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2771/3116905/f955dafd6a4d/pone.0021329.g002.jpg

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