中国动物传染病学报2026，Vol.34Issue(3)：96-105,10.
CRISPR/Cas9全基因组文库筛选RABV感染关键宿主因子方法的建立
Construction of RABV Infection-related Key Host Factor Screening Method Based on Genome-wide CRISPR/Cas9 Library
摘要
Abstract
Rabies virus(RABV)is a highly lethal neurotropic virus that infect humans and warm-blooded animals and causes rabies,which currently has no effective treatment target.In order to comprehensively and efficiently explore the key host targets that mediate RABV infection,a screening method for RABV infection-dependent host factors was established using the CRISPR/Cas9 genome-wide library.In this study,CRISPR/Cas9 genome-wide knockout lentivirus was used to infect RABV-susceptible cells HEK-293,and the lentivirus-infected cells were cultured with puromycin to obtain a genome-wide knockout cell pool.The recombinant strain CVS11-eGFP carrying the green fluorescent reporter gene was used to infect the knockout cell pool,and the cells with extremely weak green fluorescence intensity were sorted by flow cytometry 48 hours later.The gene-edited cell clones KL-4 and KL-14 that were not infected with RABV were obtained after cell monoclonal culture and secondary infection with different RABV strains.In this study,a method was established to systematically screen the key host factors that mediated RABV infection using the CRISPR/Cas9 genome-wide knockout library,which provided new technical support for the discovery of key targets for rabies treatment in the host genome.关键词
狂犬病毒/CRISPR/Cas9全基因组敲除文库/感染复制关键因子/筛选方法Key words
Rabies virus/CRISPR/Cas9 genome-wide screening library/key factors for infection and replication/screening分类
农业科技引用本文复制引用
张欣莹,郭建雄,王洁,涂长春,刘艳..CRISPR/Cas9全基因组文库筛选RABV感染关键宿主因子方法的建立[J].中国动物传染病学报,2026,34(3):96-105,10.基金项目
国家自然科学基金项目(32273093,31972720) （32273093,31972720）
国家重点研发计划(2022YFD1800100) （2022YFD1800100）
