Protection Against West Nile Virus by RNA Interference

描述（由申请人提供）：西尼罗河（WN）病毒是一种黄病毒，可引起致命的人类脑炎。在过去的四年里，这种病毒在美国各地生根发芽。目前还没有获得批准的疫苗或疗法来预防寨卡病毒。本项目旨在开发一种有效的抗wn病毒RNA干扰疗法。我们的假设是，靶向WN病毒rna降解的小干扰rna （sirna）会抑制病毒在体内的复制和传播。在培养细胞中抑制WN病毒的几种sirna将在筛选试验中被鉴定出来。在细胞检测中表现出强大干扰的sirna将在体内使用已建立的WN病毒感染模型进行评估。sirna将在腺病毒相关病毒载体上表达，包括设计用于选择性靶向WN病毒感染细胞的载体。AAV载体可在小鼠脾、肝、脑等易感器官中表达sirna。在感染WN病毒后，将评估小鼠不同器官的病毒负荷，并观察其发病率和死亡率。这些实验将为优化干扰rna递送到WN病毒感染动物模型感染器官的II期实验提供基础。

DESCRIPTION (provided by applicant): West Nile (WN) virus is a flavivirus that can cause fatal human encephalitis. Over the past four years, this virus has established itself across the United States. There are no approved vaccines or therapies to protect against WN virus. This project is designed to develop an effective anti-WN virus RNA interference therapy. Our hypothesis is that small interfering RNAs (siRNAs) targeting WN virus RNAs for degradation will inhibit viral replication and dissemination in vivo. Several siRNAs that inhibit WN virus in cultured cells will be identified in screening assays. The siRNAs showing robust interference in cellular assays will be evaluated in vivo using an established model of WN virus infection. The siRNAs will be expressed from adenovirus-associated virus vectors, including vectors designed to selectively target WN virus-infected cells. The AAV vectors will express siRNAs in mouse spleen, liver and brain, organs susceptible to WN virus. After WN virus challenge, the mice will be assessed for viral burden in different organs, and observed for morbidity, mortality. These experiments will provide the basis for Phase II experiments optimizing delivery of interfering RNAs to infected organs in animal models of WN virus infection.