Ye Yu, Zhang Mengting, Nie Haobing, Duan Yiwen, Ye Zaijiao, Zheng Chunfu
College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, China.
Jiangxi Engineering Research Center for Animal Health Products, Nanchang, Jiangxi, China.
Methods Mol Biol. 2025;2940:273-281. doi: 10.1007/978-1-0716-4615-1_24.
The whole-genome library for CRISPR screening serves as an important biotechnological tool aimed at probing gene function in mammalian cells, providing a foundation for the systematic discovery of essential genes corresponding to biological effects. Research indicates that whole-genome library cell lines can be used to identify host factors and potential drug targets associated with viral infections effectively at high throughput, providing crucial evidence for the development of novel antiviral drugs. Here, we primarily discuss the methods for constructing genome-scale CRISPR screens in cell lines and their applications in virology research. By cloning and expressing the whole genomic DNA of specific organisms, stable cell lines, which can be utilized for functional validation, drug screening, and gene function studies, can be established. Through gene knockout or overexpression techniques, in-depth analyses of the key roles that genes play in the viral life cycle could be conducted, revealing how viruses exploit the biological mechanisms of host cells for replication and evasion of immune responses. These findings not only enhance our understanding of the interactions between viruses and host cells but also yield new targets for the development of antiviral drugs and vaccines.
用于CRISPR筛选的全基因组文库是一种重要的生物技术工具,旨在探究哺乳动物细胞中的基因功能,为系统发现与生物学效应相对应的必需基因奠定基础。研究表明,全基因组文库细胞系可用于高通量有效地鉴定与病毒感染相关的宿主因子和潜在药物靶点,为新型抗病毒药物的开发提供关键证据。在此,我们主要讨论在细胞系中构建基因组规模CRISPR筛选的方法及其在病毒学研究中的应用。通过克隆和表达特定生物体的全基因组DNA,可以建立可用于功能验证、药物筛选和基因功能研究的稳定细胞系。通过基因敲除或过表达技术,可以深入分析基因在病毒生命周期中所起的关键作用,揭示病毒如何利用宿主细胞的生物学机制进行复制和逃避免疫反应。这些发现不仅增进了我们对病毒与宿主细胞之间相互作用的理解,还为抗病毒药物和疫苗的开发提供了新的靶点。