Interference to Dengue Virus Replication in Mosquitoes

描述：(申请人提供)：登革热病毒，血清1-4型为 目前影响人类的最重要的蚊媒病毒。它是 估计有25亿人面临感染的风险。大约50-100 百万例登革热病例和10万例危及生命的病例 登革出血热(DHF)每年在全球范围内发生。一个主要因素 促进了DF和DI-IF作为公共卫生在全球的戏剧性出现 令人担忧的是埃及伊蚊在整个热带地区的扩散， 登革热病毒的主要媒介。这项研究的长期目标是 确定降低或消除Ae能力的分子策略。 埃及伊蚊传播登革热和其他黄病毒。黄病毒在中国复制 蚊虫细胞内双链表达可抑制蚊虫细胞 核糖核酸。RNA介导的干扰蚊子细胞的特点是 类似于植物进化上保守的防御机制 其他无脊椎动物，如果蝇。我们假设 在蚊子中，RNA介导的干扰(RMI)是一个强大的细胞过程， 与其他生物体中的那些同源，并且可以被操纵来 消除蚊子对黄病毒的媒介能力。在这项提案中，我们 计划进行研究，以确定蚊子RMI的详细机制。在……里面 除了我们的机制研究之外，我们还将检查RMI在 在蚊子和蚊子中维持黄病毒持续感染 确定黄病毒传播的媒介能力。 该建议的具体目的如下：1.明确 RNA介导的干扰(RMI)--对蚊子中黄病毒的干扰。分子 以及培养的蚊虫细胞和Ac中RMI的生化特性。 埃及伊蚊将与转录后基因沉默(Ptgs)进行比较。 在植物和果蝇中的RNA干扰(RNAi)。2：确定 蚊虫持续虫媒病毒感染与RNAi的关系 RMI的反防御性登革热病毒抑制子可能是 通过使用登革热病毒感染性克隆进行检查。3.确定 RNAi在蚊子对黄病毒的耐受性/敏感性中的作用交流。 与RMI反应相关的埃及伊蚊基因将被表征并 已映射。

DESCRIPTION: (provided by the applicant): Dengue viruses, serotypes 1-4 are currently the most important mosquito-borne viruses affecting humans. It is estimated that 2.5 billion humans are at risk for infection. About 50-100 million cases of dengue fever (DF) and >100,000 cases of life-threatening dengue hemorrhagic fever (DHF) occur annually worldwide. A major factor contributing to the dramatic global emergence of DF and DI-IF as public health concerns is the proliferation throughout the tropics of Aedes aegypti, the principal vector of DEN viruses. The long-term goal of this research is to define molecular strategies for reducing or eliminating the competence of Ae. aegypti to transmit DEN and other flaviviruses. Flavivirus replication in mosquito cells can be inhibited by intracellular expression of double-stranded RNA. Characteristics of RNA-mediated interference in mosquito cells are very similar to those of evolutionarily conserved defense mechanisms in plants and other invertebrates such as Drosophila melanogaster. We hypothesize that RNA-mediated interference (RMI) in mosquitoes is a robust cellular process, homologous to those in other organisms, and that it can be manipulated to eliminate mosquito vector competence for flaviviruses. In this proposal, we plan studies to determine the detailed mechanism of RMI in mosquitoes. In addition to our mechanistic studies, we will examine the role of RMI in maintenance of flavivirus persistent infections in mosquitoes and in determining vector competence for flavivirus transmission. The specific aims of the proposal are as follows: 1.To define the mechanism for RNA-mediated interference (RMI)-to flaviviruses in mosquitoes. The molecular and biochemical characteristics of RMI in cultured mosquito cells and Ac. aegypti will be compared to those of posttranscriptional gene silencing (PTGS) in plants and RNA interference (RNAi) in Drosophila. 2:To determine the relationship between RNAi and persistent arbovirus infections in mosquitoes. The possibility that a counterdefensive dengue viral suppressor of RMI will be examined by use of a dengue virus infectious clone. 3. To determine the role of RNAi in refractoriness/susceptibility of mosquitoes to flaviviruses. Ac. aegypti genes associated with the RMI response will be characterized and mapped.