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利用 CRISPR 筛选技术进行病毒宿主因子发现的研究综述

A review of virus host factor discovery using CRISPR screening.

机构信息

Program in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore, Singapore.

Infectious Diseases Labs (A*STAR ID Labs), Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.

出版信息

mBio. 2024 Nov 13;15(11):e0320523. doi: 10.1128/mbio.03205-23. Epub 2024 Oct 18.

Abstract

The emergence of genome-scale forward genetic screening techniques, such as Haploid Genetic screen and clustered regularly interspaced short palindromic repeats (CRISPR) knockout screen has opened new horizons in our understanding of virus infection biology. CRISPR screening has become a popular tool for the discovery of novel host factors for several viruses due to its specificity and efficiency in genome editing. Here, we review how CRISPR screening has revolutionized our understanding of virus-host interactions from scientific and technological viewpoints. A summary of the published screens conducted thus far to uncover virus host factors is presented, highlighting their experimental design and significant findings. We will outline relevant methods for customizing the CRISPR screening process to answer more specific hypotheses and compile a glossary of conducted CRISPR screens to show their design aspects. Furthermore, using flaviviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as examples, we hope to offer a broad-based perspective on the capabilities of CRISPR screening to serve as a reference point to guide future unbiased discovery of virus host factors.

摘要

基因组规模正向遗传筛选技术的出现,如单倍体遗传筛选和成簇规律间隔短回文重复(CRISPR)敲除筛选,为我们理解病毒感染生物学开辟了新的视野。CRISPR 筛选因其在基因组编辑中的特异性和效率,已成为发现几种病毒新宿主因子的流行工具。在这里,我们从科学和技术的角度回顾了 CRISPR 筛选如何彻底改变我们对病毒-宿主相互作用的理解。总结了迄今为止为发现病毒宿主因子而进行的已发表的筛选,突出了它们的实验设计和重要发现。我们将概述定制 CRISPR 筛选过程以回答更具体假设的相关方法,并编制一个已进行的 CRISPR 筛选词汇表,以展示其设计方面。此外,我们还希望以黄病毒和严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)为例,提供一个广泛的视角,展示 CRISPR 筛选作为一个基准点,以指导未来对病毒宿主因子的无偏发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ba4/11559068/d26670e6cd19/mbio.03205-23.f001.jpg

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