中国兽医科学2025，Vol.55Issue(12)：1599-1607,9.DOI:10.16656/j.issn.1673-4696.2025.0216
敲除ATG5基因Vero-SN细胞系的建立
Establishment of Vero-SN cell line with ATG5 gene knockout
摘要
Abstract
To explore how viruses exploit host cellular autophagy mechanisms for replication and dissemination and to provide a research model,this study utilized the CRISPR/Cas9 system to establish ATG5 gene knockout cell lines.Initially,the sgRNA sequences targeting ATG5 gene in African green monkey kidney cells(Vero)were designed and cloned into lentiviral vector,the re-packaged lentivirus was then used to infect the Vero-SN cells,and then screened to obtain ATG5 gene knockout subclone cell lines.The replication ability of peste des petits ruminants virus(PPRV)in Vero-SN cells and ATG5 gene knockout cell lines was detected using TCID50 and Western-blot,respectively.The results showed that the VeroSN-ATG5KO cell line was successfully established,and the inhibition of autophagy significantly impaired viral replication.Establishing this gene-edited cell line provides crucial experimental material for further investigation of the interaction and molecular mechanisms between viruses and autophagic proteins.关键词
CRISPR/Cas9/Vero-SN细胞系/ATG5基因/自噬/基因敲除Key words
CRISPR/Cas9/Vero-SN cell line/ATG5 gene/autophagy/gene knockout分类
生物科学引用本文复制引用
刘迪一,陈炳春,杨雪,王会宝,殷相平,任善会,窦永喜..敲除ATG5基因Vero-SN细胞系的建立[J].中国兽医科学,2025,55(12):1599-1607,9.基金项目
国家现代农业产业技术体系建设专项(CARS-39-13) （CARS-39-13）
甘肃省科技重大专项(23ZDNA007) （23ZDNA007）
甘肃省自然科学基金项目(24JRRE011) （24JRRE011）
高校教师创新基金项目(2025A-326) （2025A-326）
