Yang Yang, Zheng Jiayang, Lin Yafang, Zhang Yan, Zhao Qianming, Zhang Hailong, Zhang Junjie, Li Zongjie, Liu Ke, Li Beibei, Shao Donghua, Qiu Yafeng, Ma Zhiyong, Wei Jianchao
Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Viruses. 2025 May 27;17(6):759. doi: 10.3390/v17060759.
Viral replicons are efficient tools to understand the mechanisms of viral replication and screen antiviral drugs. In this study, a viral-cDNA-based replicon of Japanese encephalitis virus (JEV), which is the causative agent of Japanese encephalitis, was constructed by replacing the viral structural proteins with a green fluorescent protein (JEV-GFP replicon). The resulting JEV-GFP replicon was used as a tool to screen antiviral drugs targeting JEV nonstructural proteins, and the five compounds JNJ-A07, HZ-1157, NITD-2, quinine, and NITD008 were obtained, which significantly inhibited the replication of the JEV-GFP replicon and JEV in vitro, and the properties of these five compounds were also analyzed. The CC, EC, and SI indices of these five compounds were analyzed. In addition, the JEV-GFP replicon was used as a tool to identify the residues of viral nonstructural proteins involved in RNA replication, and the cysteine residue at position 4 of nonstructural protein 1 was found to be essential for JEV RNA replication. These data suggested that the noninfectious JEV-GFP replicon could be used as tool for different purposes, such as antiviral drug screening and gene function studies.
病毒复制子是了解病毒复制机制和筛选抗病毒药物的有效工具。在本研究中，通过用绿色荧光蛋白替换病毒结构蛋白构建了一种基于病毒cDNA的日本脑炎病毒（JEV，日本脑炎的病原体）复制子（JEV-GFP复制子）。所得的JEV-GFP复制子被用作筛选靶向JEV非结构蛋白的抗病毒药物的工具，并获得了JNJ-A07、HZ-1157、NITD-2、奎宁和NITD008这五种化合物，它们在体外显著抑制了JEV-GFP复制子和JEV的复制，并且还分析了这五种化合物的性质。分析了这五种化合物的CC、EC和SI指数。此外，JEV-GFP复制子被用作鉴定参与RNA复制的病毒非结构蛋白残基的工具，发现非结构蛋白1第4位的半胱氨酸残基对JEV RNA复制至关重要。这些数据表明，非感染性的JEV-GFP复制子可作为用于不同目的的工具，如抗病毒药物筛选和基因功能研究。
