锌指蛋白33B(Zinc finger protein 33B, ZNF33B)抑制乙型脑炎病毒复制的功能与分子机制研究

乙型脑炎病毒(JEV)能突破血脑屏障导致脑部炎症和神经损伤，是一种重要的人兽共患传染病病原。目前尚无有效药物和治疗制剂，因此加强JEV与宿主互作的研究，探寻有效抗病毒靶点和治疗方法尤为重要。本课题组前期通过CRISPR/Cas9技术筛选到锌指蛋白33B(ZNF33B)影响JEV增殖，但其作用机制及生物学效应尚不清楚。本研究拟以猪ZNF33B抑制JEV增殖为切入点，利用病毒学、免疫学和细胞生物学等方法进行（1）猪ZNF33B对JEV复制的抑制效果；（2）猪ZNF33B与JEV病毒RNA互作的分子细节和翻译的影响以及生物学效应；（3）猪ZNF33B相互作用的宿主蛋白筛选及协同抗病毒效应；（4）猪ZNF33B在JEV抗病毒天然免疫中的作用等的研究，阐明其抑制JEV复制的分子机制，为JEV抗病毒药物设计和分子抗育种研究提供理论依据，具有重要的理论和实践意义。

Japanese encephalitis virus (JEV) is a mosquito-borne zoonotic virus that targets the central nervous system (CNS) and cause brain inflammation and nerve damage with high morbidity and mortality. Currently, there are no effective drugs and therapeutic agents approved for the treatment of JEV infection. The discovery of disease resistance genes and the elucidation of their molecular mechanism are keys to molecular disease resistance breeding. Our preliminary data showed that knockdown of porcine zinc finger protein 33B(ZNF33B) facilitates JEV propagation, which showed that ZNF33B take on antiviral activity to JEV. However, the molecular mechanisms and biological effects of porcine ZNF33B during JEV infection remain elusive. In this project, we propose to determine the inhibition effect of porcine ZNF33B against JEV and clarity the underlying mechanism. We will carry out the following studies from two aspects of virus and host by using virology, immunology and cell biology methods. 1) furtherly evaluated its abilities of inhibition activity against JEV replication by overexpressing and knock-out porcine ZNF33B protein; 2) detect the ability of porcine ZNF33B binds to JEV viral RNA, the influence on viral translation and viral RNA stability; 3) Identify the host factors interacted with porcine ZNF33B through immunoprecipitation combined with mass spectrometry analysis and investigate the potential cooperative action or antagonism role(s) with porcine ZNF33B; 4) Reveal the role of ZNF33B in the antiviral natural immunity during JEV infection. Completion of this project will be useful to clarity the antiviral function and molecular mechanism of porcine ZNF33B against JEV, and provides novel insights into the development of novel vaccine, antiviral drug, and generation of genetic resistance animals for preventing the dissemination of the infectious disease in the future.