Arboviral suppressors of RNA interference

描述（由申请人提供）：黄热病蚊子，埃及伊蚊，是人类疾病的多产媒介。我们的长期目标是在分子水平上了解蚊子体内病毒-载体相互作用的基础上，开发控制节肢动物传播病毒的新方法。已经很好地证明节肢动物传播的病毒对蚊子的RNAi反应敏感。因此，RNA干扰（RNAi）可能在媒介蚊子中存在的几种潜在病原体传播屏障中发挥作用。要评估的假设是节肢动物传播的病毒已经进化到在其基因组中编码RNAi反应的抑制因子。本提案的具体目的是：(1)利用稳定的种系转化来建立和验证Ae。埃及伊蚊“RNAi传感器”菌株。(2)利用产生的传感器菌株检测候选节肢动物传播的病毒是否存在沉默抑制因子。这些目标的完成也将为研究病毒-载体相互作用、蚊子遗传学和病毒遗传学产生一种新的分析系统。与蚊媒病毒引起的疾病相关的总发病率、死亡率和经济损失是不可估量的。有证据表明，RNA沉默可能是对蚊子体内病毒积累的一种普遍限制。鉴定蚊子传播的病毒基因组中编码的RNA沉默抑制子将对了解节肢动物传播的病毒性疾病的传播具有巨大的意义。

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.