Wang Jing, Zheng Chunfu, Xiao Wuhan
Key Laboratory of Breeding Biotechnology and Sustainable Aquaculture, Institute of Hydrobiology, Chinese Academy of Sciences, Hubei Hongshan Laboratory, Wuhan, China.
University of Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2025;2940:261-272. doi: 10.1007/978-1-0716-4615-1_23.
During the long evolutionary process, viruses and hosts have engaged in a game of immune escape and viral clearance, resulting in an exquisite model of virus-host interaction. An appropriate animal model can help unravel the mysteries of this process. Owing to their susceptibility to multiple viruses, small size, high fecundity, low cost, ease of visualization, and efficient gene editing, zebrafish (Danio rerio) have recently emerged as a powerful vertebrate model system for studying virus-host interactions. In this chapter, we describe in detail the experimental protocols for analyzing virus-host interactions in a zebrafish model in which spring viremia of carp virus (SVCV) is used as an example. These protocols include the generation of knockout zebrafish via CRISPR/Cas9 technology, methods for viral infection of zebrafish larvae or adults, analysis of in vivo viral resistance of host genes, analysis of host and viral gene expression in zebrafish larvae via qRT-PCR and Western blotting, and detection of host protein binding to viral proteins via immunoprecipitation. These experimental protocols provide an effective reference for studying virus-host interactions in a zebrafish model.
在漫长的进化过程中，病毒与宿主展开了一场免疫逃逸与病毒清除的较量，形成了一种精妙的病毒-宿主相互作用模式。合适的动物模型有助于揭开这一过程的奥秘。由于斑马鱼（Danio rerio）对多种病毒易感、体型小、繁殖力高、成本低、易于观察且基因编辑效率高，它们最近已成为研究病毒-宿主相互作用的强大脊椎动物模型系统。在本章中，我们详细描述了以鲤春病毒血症病毒（SVCV）为例，在斑马鱼模型中分析病毒-宿主相互作用的实验方案。这些方案包括通过CRISPR/Cas9技术构建基因敲除斑马鱼、斑马鱼幼体或成体的病毒感染方法、宿主基因体内抗病毒抗性分析、通过qRT-PCR和蛋白质免疫印迹分析斑马鱼幼体中宿主和病毒基因表达，以及通过免疫沉淀检测宿主蛋白与病毒蛋白的结合。这些实验方案为在斑马鱼模型中研究病毒-宿主相互作用提供了有效的参考。
