中国兽医科学2025，Vol.55Issue(6)：725-731,7.DOI:10.16656/j.issn.1673-4696.2025.0097
CRISPR-Cas9介导的TRIM3基因敲除细胞系的建立及其应用
Establishment and application of TRIM3 gene knockout cell line mediated by CRISPR-Cas9
摘要
Abstract
This research project is dedicated to exploring the influence of the host protein TRIM3 on the replication of foot-and-mouth disease virus(FMDV).Initially,the CRISPR/Cas9 system was employed to establish the TRIM3 gene knockout BHK21 cell line.The effect of this gene's deficiency on cell viability was examined with the CCK8 kit.The results indicated that knockout of TRIM3 had no impact on cell growth.Subsequently,wild-type and TRIM3 knockout BHK21 cells were infected with FMDV separately.The replication of FMDV in the above two cell lines was detected through Q-PCR,Western-blot and TCID50.The results indicated that the absence of TRIM3 could inhibit the replication of FMDV.To further verify the phenotypic function of TRIM3,a cell line overexpressing TRIM3 was constructed using a lentiviral packaging system.The results showed that overexpression of TRIM3 could promote the replication of FMDV.In conclusion,this study constructed the TRIM3 gene knockout and overexpression cell lines and proved for the first time that TRIM3 can positively regulate the replication of FMDV,laying a foundation for the subsequent exploration of the specific mechanisms by which TRIM3 regulates the replication of FMDV.关键词
TRIM3/口蹄疫病毒/细胞系Key words
TRIM3/foot-and-mouth disease virus(FMDV)/cell lines分类
畜牧业引用本文复制引用
裴丹诗,万博,范许许,朱兆宇,王一卓,何路,任青峰,李熙忠,郑海学,李伟伟..CRISPR-Cas9介导的TRIM3基因敲除细胞系的建立及其应用[J].中国兽医科学,2025,55(6):725-731,7.基金项目
国家自然科学基金青年项目(32202781,32302857) （32202781,32302857）
国家自然科学基金重点项目(32330107) （32330107）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
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项目(1610312021009) （1610312021009）
