College of Medicine and Public Health, Flinders University, Bedford Park, SA 5042, Australia.
Viruses. 2021 May 28;13(6):1020. doi: 10.3390/v13061020.
Several recently developed high-throughput techniques have changed the field of molecular virology. For example, proteomics studies reveal complete interactomes of a viral protein, genome-wide CRISPR knockout and activation screens probe the importance of every single human gene in aiding or fighting a virus, and ChIP-seq experiments reveal genome-wide epigenetic changes in response to infection. Deep mutational scanning is a relatively novel form of protein science which allows the in-depth functional analysis of every nucleotide within a viral gene or genome, revealing regions of importance, flexibility, and mutational potential. In this review, we discuss the application of this technique to RNA viruses including members of the Flaviviridae family, Influenza A Virus and Severe Acute Respiratory Syndrome Coronavirus 2. We also briefly discuss the reverse genetics systems which allow for analysis of viral replication cycles, next-generation sequencing technologies and the bioinformatics tools that facilitate this research.
几种最近开发的高通量技术改变了分子病毒学领域。例如，蛋白质组学研究揭示了病毒蛋白的完整互作组，全基因组 CRISPR 敲除和激活筛选研究了人类每个基因在帮助或对抗病毒方面的重要性，而 ChIP-seq 实验揭示了感染后全基因组的表观遗传变化。深度突变扫描是一种相对较新的蛋白质科学形式，它允许对病毒基因或基因组中的每个核苷酸进行深入的功能分析，揭示重要性、灵活性和突变潜力的区域。在这篇综述中，我们讨论了该技术在 RNA 病毒中的应用，包括黄病毒科的成员、甲型流感病毒和严重急性呼吸综合征冠状病毒 2。我们还简要讨论了允许分析病毒复制周期的反向遗传学系统、下一代测序技术和促进这项研究的生物信息学工具。
