中国兽医科学2025，Vol.55Issue(10)：1291-1298,8.DOI:10.16656/j.issn.1673-4696.2025.0172
EphA2基因敲除和过表达BHK-21细胞系的构建及其对口蹄疫病毒复制的影响
Construction of EphA2 gene knockout and overexpression BHK-21 cell lines and their effects on foot-and-mouth disease virus replication
摘要
Abstract
The aim of this study was to construct erythropoietin-producing hepatocellular receptor A2(EphA2)gene knockout and overexpression BHK-21 cell lines to investigate their effects on foot-and-mouth disease virus(FMDV)replication.The EphA2 gene knockoout BHK-21 cell line was constructed by CRISPR/Cas9 gene editing technology,and the EphA2 gene overexpression BHK-21 cell line was constructed using lentivirus infection.After puromycin screening and monoclonal picking,stable cell lines were suc-cessfully obtained.Subsequently,wild-type BHK-21 cells were used as the control,and FMDV was used to infect the above cell lines respectively,and the viral nucleic acid and protein levels were detected by real-time fluorescence quantitative PCR and immunoblotting respectively.The results showed that the replication of FMDV in BHK-21 cells was significantly inhibited by EphA2 knockout,and the expression of viral nucleic acids and proteins was significantly reduced;while FMDV replication was enhanced when EphA2 gene was overexpressed.This study reveals that EphA2 gene has an important role in the replica-tion process of FMDV,which provides a theoretical basis for an in-depth understanding of the infection mechanism and prevention and control strategies of FMDV.关键词
CRISPR/Cas9/EphA2/口蹄疫病毒Key words
CRISPR/Cas9/EphA2/foot-and-mouth disease virus分类
农业科技引用本文复制引用
李熙忠,郜原,范许许,朱兆宇,裴丹诗,王一卓,何路,任青峰,郑海学,李伟伟..EphA2基因敲除和过表达BHK-21细胞系的构建及其对口蹄疫病毒复制的影响[J].中国兽医科学,2025,55(10):1291-1298,8.基金项目
国家自然科学基金青年项目(32202781,32302857) （32202781,32302857）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
国家自然科学基金重点项目(32330107) （32330107）
中国博士后科学基金项目(2023M743830) （2023M743830）
国家生猪产业技术体系项目(CARS-35,CARS-39-13) （CARS-35,CARS-39-13）
国家生猪技术创新中心先导科技项目(NCTIP-XD/C03) （NCTIP-XD/C03）
中国农业科学院创新计划项目(CAAS-CSLPDCP-2023002,CAAS-ASTIP-2023-LVRI) （CAAS-CSLPDCP-2023002,CAAS-ASTIP-2023-LVRI）
"十四五"广东省农业科技创新十大主攻方向"揭榜挂帅"项目(2024KJ14) （2024KJ14）
兰州兽医研究所基本科研业务费项目(161031202-1009) （161031202-1009）
