Aedes antiviral RNAi pathway

摘要 虫媒病毒仍然是一个巨大的公共卫生威胁，每年都会造成大规模流行病。最重要的目标 本研究的目的是更好地了解埃及伊蚊RNA干扰(RNAi)途径在 抗病毒防御和先天免疫，并产生知识和工具来开发新的 控制虫媒病毒传播的方法。野生型的转基因种群置换 不能传播病原体的蚊子正在成为其他疾病的有希望的补充 控制方法。因为Ae.埃及伊蚊传播多种经常共处的虫媒病毒病原体，它 对转基因蚊子来说，对多种病原体具有抵抗力很重要。SiRNA途径介导的 众所周知，抗病毒防御系统可以抵御广泛的病毒。然而，尽管RNAi PATH作为主要的泛抗病毒防御系统的出现在蚊子中仍未得到充分的研究。我们的 研究计划旨在检验埃及伊蚊的主要假设，即埃及伊蚊的蛋白，DICER-2(DCR-2)，R2D2 和ArgAerte-2(AGO-2)是介导具有广泛保护性的RNA沉默反应的关键成分 对抗虫媒病毒感染。我们将使用siRNA缺陷的功能丧失突变体和转基因 蚊子过度表达将在AIM 1中产生的DCR-2、AGO-2和R2D2。使用这些遗传工具 我们将在目标2中阐明抗病毒反应的时间和空间特异性，并研究可能的 目标3中的组织间信号和垂直传输的调节。在目标4中，我们将解决相互作用 在RNAi途径和其他先天免疫防御系统之间。这个项目利用了 DRS Dimopoulos和Myles与虫媒病毒感染系统、蚊子的互补专业知识 转基因、蚊子先天免疫和RNAi/小RNA生物学。我们提议的项目还将产生 为研究Ae中RNAi途径的其他方面提供了强有力的工具。埃及生物学。

SUMMARY Arboviruses remain an immense public health threat, causing yearly large epidemics. The overarching aim of this research is to better understand the role of the Aedes aegypti RNA interference (RNAi) pathway in antiviral defense and innate immunity, and to generate knowledge and tools for the development of new methods to control arbovirus transmission. Population replacement of wild-type with genetically modified mosquitoes incapable of pathogen transmission is emerging as a promising complement to other disease control methods. Because Ae. aegypti transmits multiple arbovirus pathogens that are frequently sympatric, it is important for a transgenic mosquito to be resistant to multiple pathogens. The siRNA pathway-mediated antiviral defense system is known to act against a broad range of viruses. However, despite the RNAi pathway's emergence as the major pan-antiviral defense system it remains understudied in mosquitoes. Our research plan is designed to test the overarching hypothesis that the A. aegypti proteins, Dicer-2 (Dcr-2), R2D2 and Argonaute-2 (Ago-2), are key components mediating an RNA silencing response that is broadly protective against arbovirus infections. We will use of siRNA-deficient loss-of-function mutants and transgenic mosquitoes over-expressing Dcr-2, Ago-2 and R2D2 that will be generated in Aim 1. With these genetic tools we will clarify the temporal and spatial specificity of the antiviral response in Aim 2, and investigate possible inter-tissue signaling and regulation of vertical transmission in Aim 3. In Aim 4 we will address interactions between the RNAi pathway and other innate immunity defense systems. This project utilizes the complementary expertise of Drs Dimopoulos and Myles with the arbovirus infection systems, mosquito transgenesis, mosquito innate immunity and RNAi/small RNA biology. Our proposed project will also generate powerful tools for studying other aspects of the RNAi pathway in Ae. aegypti biology.