中国兽医科学2024，Vol.54Issue(8)：1010-1018,9.DOI:10.16656/j.issn.1673-4696.2024.0173
METTL3表达水平对口蹄疫病毒复制的影响
Effect of METTL3 expression level on foot-and-mouth disease virus replication
摘要
Abstract
METTL3 gene knockout cell line(BHK-21)was firstly constructed utilizing CRISPR/Cas9 system.Subsequently,wild-type and METTL3 knockout cells were infected with foot-and-mouth disease virus(FMDV),and the replication of FMDV in these two cells was detected by qPCR,Western-blot and TCID50 experiments.The results indicated that METTL3 deficiency significantly inhibited FMDV replication.To further verify the function of METTL3,a stable METTL3 expression cell line was constructed using the lentivirus packaging system.Wild-type and METTL3 overexpressed cells were infected with FMDV,and the replication of FMDV in these two cells was also measured by qPCR and TCID50 assays.The results showed that overexpression of METTL3 promoted FMDV replication.In addition,laser confocal microscopy found that METTL3 migrated from the nucleus to cytoplasm upon FMDV infection,and the change of intracellular localization suggested that METTL3 might be involved in regulating the transcription process of FMDV genome.In this study,we successfully constructed a BHK-21 cell line with METTL3 knockout and overex-pression,and found that METTL3 positively regulates FMDV replication.These results provide a reliable data base for further exploration on the mechanism of METTL3 regulation of FMDV replication in BHK-21 cells.关键词
METTL3/m6A修饰/口蹄疫病毒Key words
methyltransferase-like 3/N6-methyladenosine modification/foot-and-mouth disease virus分类
农业科技引用本文复制引用
张继燕,任青峰,张伟,王川,郑海学,李伟伟,范许许,曹雪静,朱兆宇,裴丹诗,王一卓,何路,李熙忠..METTL3表达水平对口蹄疫病毒复制的影响[J].中国兽医科学,2024,54(8):1010-1018,9.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(2021YFD1801300,2021YFD1800300) （2021YFD1801300,2021YFD1800300）
甘肃省青年科技基金计划项目(22JR5RA034) （22JR5RA034）
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(2023SDZG02) （2023SDZG02）
兰州兽医研究所基本科研业务费项目(1610312021009) （1610312021009）
