Arboviral suppressors of RNA interference

描述(申请人提供)：黄热病蚊子埃及伊蚊是人类疾病的多发媒介。我们的长期目标是开发新的方法来控制节肢动物传播的病毒，基础是在分子水平上了解蚊子中发生的病毒与媒介的相互作用。已经很好地证明了节肢动物传播的病毒对蚊子中发现的RNAi反应敏感。因此，RNA干扰(RNAi)可能在媒介蚊子内存在的几个潜在的病原体传播障碍中发挥作用。需要评估的假设是，节肢动物传播的病毒已经进化到编码它们基因组中的RNAi反应抑制因子。这一建议的具体目的是：(1)使用稳定的生殖系转化来建立和验证Ae。埃及伊蚊“RNAi传感器”菌株。(2)使用产生的传感器菌株来检测候选节肢动物传播的病毒是否存在沉默抑制子。完成这些目标还将产生一种新的检测系统，用于研究病毒-媒介相互作用、蚊子遗传学和病毒遗传学。蚊媒病毒引起的疾病的总发病率、死亡率和经济损失是不可估量的。有证据表明，RNA沉默可能代表了对蚊子体内病毒积累的普遍限制。识别蚊媒病毒基因组中编码的RNA沉默抑制因子将对了解节肢动物传播病毒疾病具有巨大的意义。

DESCRIPTION (provided by applicant): The yellow fever mosquito, Aedes aegypti, is a prolific vector of human disease. Our long-term goal is to develop new methods for controlling arthropod-borne viruses based on understanding at the molecular level the virus-vector interactions occurring in the mosquito. It has been well demonstrated that arthropod-borne viruses are susceptible to the RNAi responses found in mosquitoes. Thus, RNA interference (RNAi) may play a role in several potential barriers to pathogen transmission present within the vector mosquito. The HYPOTHESIS to be evaluated is that arthropod-borne viruses have evolved to encode suppressors of the RNAi response within their genomes. The specific aims of this proposal are to: (1) Use stable germline transformation to establish and validate an Ae. aegypti "RNAi sensor" strain. (2) Use the sensor strain generated to assay candidate arthropod-borne viruses for the presence of silencing suppressors. Completion of these aims will also generate a novel assay system for the study virus-vector interactions, mosquito genetics, and virus genetics. The total morbidity, mortality, and economic loss associated with diseases caused by mosquito-borne viruses are inestimable. Evidence indicates that RNA silencing may represent a general constraint to the accumulation of virus in the mosquito. The identification of suppressors of RNA silencing encoded in mosquito-borne virus genomes would have enormous implications for understanding the transmission of arthropod-borne viral diseases.