基于CRISPR/Cas9技术的高通量筛选平台:发掘病毒复制相关宿主分子的新途径

病毒作为严格的细胞内寄生生物,需要多种宿主蛋白辅助其完成生命周期。寻找与病毒复制相关的宿主因子并揭示其作用机制,将有助于阐明病毒的感染机制,为疫病的防治提供新靶标。与RNA干扰技术相比,近年来兴起的CRISPR/Cas9技术能更特异、高效、准确地实现基因组编辑,因而在功能基因研究中得到更广泛应用。而基于CRISPR/Cas9系统的宿主全基因组sgRNA文库高通量筛选技术平台,可快速发现参与病毒侵入、复制等生物学过程的关键宿主因子,通过明确病毒-宿主分子相互作用进而揭示病毒的生命周期,为分子病毒学和免疫学提供了强大的研究工具。本文主要总结了基于CRISPR/Cas9技术的高通量筛选平台的具体筛选流程,归纳和讨论了该平台在筛选调控病毒复制相关宿主因子中的应用现状和发展前景。

CRISPR/Cas9-based high-throughput screen platform: an emerging powerful tool for the discovery of host factors involved in viral replication

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.