Hu Shijie, Gan Mailin, Wei Ziang, Shang Pan, Song Lei, Feng Jinkang, Chen Lei, Niu Lili, Wang Yan, Zhang Shunhua, Shen Linyuan, Zhu Li, Zhao Ye
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan, Sichuan Agricultural University, Ya'an, China.
State Key Laboratory of Swine and Poultry Breeding Industry, Sichuan Agricultural University, Ya'an, China.
Front Microbiol. 2024 Nov 21;15:1498641. doi: 10.3389/fmicb.2024.1498641. eCollection 2024.
Genome-wide CRISPR library screening technology is a gene function research tool developed based on the CRISPR/Cas9 gene-editing system. The clustered regularly interspaced short palindromic repeats/CRISPR-associated genes (CRISPR/Cas) system, considered the third generation of gene editing after zinc finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN), is widely used for screening various viral host factors. CRISPR libraries are classified into three main categories based on the different functions of Cas9 enzymes: CRISPR knockout (CRISPR KO) library screening, CRISPR transcriptional activation (CRISPRa) library screening, and CRISPR transcriptional interference (CRISPRi) library screening. Recently, genome-wide CRISPR library screening technology has been used to identify host factors that interact with viruses at various stages, including adsorption, endocytosis, and replication. By specifically modulating the expression of these host factors, it becomes possible to cultivate disease-resistant varieties, establish disease models, and design and develop vaccines, among other applications. This review provides an overview of the development and technical processes of genome-wide CRISPR library screening, as well as its applications in identifying viral host factors in livestock and poultry.
全基因组CRISPR文库筛选技术是一种基于CRISPR/Cas9基因编辑系统开发的基因功能研究工具。成簇规律间隔短回文重复序列/CRISPR相关基因（CRISPR/Cas）系统被认为是继锌指核酸酶（ZFN）和转录激活样效应核酸酶（TALEN）之后的第三代基因编辑技术，广泛应用于筛选各种病毒宿主因子。根据Cas9酶的不同功能，CRISPR文库主要分为三类：CRISPR敲除（CRISPR KO）文库筛选、CRISPR转录激活（CRISPRa）文库筛选和CRISPR转录干扰（CRISPRi）文库筛选。近年来，全基因组CRISPR文库筛选技术已被用于鉴定在吸附、内吞和复制等各个阶段与病毒相互作用的宿主因子。通过特异性调节这些宿主因子的表达，可以培育抗病品种、建立疾病模型以及设计和开发疫苗等。本文综述了全基因组CRISPR文库筛选的发展和技术流程，以及其在鉴定畜禽病毒宿主因子方面的应用。
