Abstract:Virus, as a strict intracellular parasitic organism, needs to use a variety of host proteins to complete their life cycle. To prevent and control viral diseases, it is necessary to provide useful data for understanding the replication and pathogenesis of viruses. Compared with the traditional RNA interference screening, CRISPR/Cas9 technology, as an emerging approach in recent years, exhibits more specific and faster characterization to achieve gene knockout or other genome editing. Therefore, it has wide applications in the study of functional genes. Based on the CRISPR/Cas9 system, we can construct a host genome-wide sgRNA library and further screen and identify the key host factors involved in multiple biological processes of pathogen invasion or replication. Lastly, we hope that the virus life cycle will be completely revealed in terms of virus-host interaction network, and high-throughput screening platform based on CRISPR/Cas9 technology will provide a powerful tool for researching virology and immunology. This review aims to introduce the screening steps of CRISPR/Cas9-based high-throughput screening platform, and summarize the applications and outlooks of this platform in screening host factors involved in viral replication.