中国兽医科学2025，Vol.55Issue(6)：711-718,8.DOI:10.16656/j.issn.1673-4696.2025.0109
LULL1基因敲除或过表达PK-15细胞系的构建及其对口蹄疫病毒复制的影响
Construction of LULL1 gene knockout or overexpression PK-15 cell line and its effect on foot-and-mouth disease virus replication
摘要
Abstract
The purpose of this study was to explore the effect of host factor LULL1 on foot-and-mouth disease virus(FMDV)replication,and to provide new insights into the mechanism of virus-host interaction.Firstly,the LULL1 gene knockout cell line PK-15-LULL1-KO was constructed by CRISPR/Cas9 gene editing technology,and then the cell viability was detected by CCK8 kit,and it was found that LULL1 gene knockout had no significant effect on cell viability,and the effects of PK-15-LULL1-KO and PK-15 wild-type on FMDV replication were compared,and the transcriptional level of FMDV mRNA was detected by real-time quantitative PCR.The expression of FMDV nonstructural protein 2C was detected by Western-blot,and the viral titer changes in the supernatant were detected by TCID50,and the results showed that knockout of LULL1 gene could significantly inhibit FMDV replication.In order to further explore the phenotypic function of LULL1,a lentiviral packaging system was used to construct a LULL1-overexpressing PK-15 cell line,which significantly promoted the replication of FMDV compared with PK-15 wild-type cells.In conclusion,LULL1 gene knockout and overexpression cell lines were successfully constructed,which will provide a theoretical basis for further elucidating the mechanism of LULL1 promoting FMDV replication and regulating FMDV replication.关键词
口蹄疫病毒/LULL1/细胞系/非结构蛋白2CKey words
foot-and-mouth disease virus/LULL1/cell line/nonstructural protein 2C分类
畜牧业引用本文复制引用
王一卓,李伟伟,蒋大伟,范许许,朱兆宇,裴丹诗,何路,李熙忠,任青峰,郑海学,朱紫祥..LULL1基因敲除或过表达PK-15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(6):711-718,8.基金项目
国家重点研发计划项目(2021YFD1800302) （2021YFD1800302）
国家自然科学基金青年项目(32302857,32202781) （32302857,32202781）
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）
吉林大学人兽共患病研究教育部重点实验室资助项目(2023iz001) （2023iz001）
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(1610312021009) （1610312021009）
