首页|METTL3表达水平对口蹄疫病毒复制的影响

METTL3表达水平对口蹄疫病毒复制的影响

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
利用CRISPR/Cas9系统构建了 METTL3基因敲除细胞系(BHK-21),随后用口蹄疫病毒(FMDV)分别感染野生型和METTL3敲除BHK-21细胞,通过qPCR、Western-blot和TCID50方法检测FMDV在上述2株细胞中的复制情况.结果表明,敲除METTL3显著抑制FMDV的复制.为了进一步验证METTL3的表型功能,利用慢病毒包装系统构建METTL3稳定表达细胞系.用FMDV分别感染野生型和METTL3过表达BHK-21细胞,通过qPCR和TCID50试验测定FMDV在这2株细胞中的复制情况.结果显示,过量表达METTL3促进FMDV的复制.此外,激光共聚焦显微镜试验显示,FMDV感染BHK-21细胞后,METTL3从细胞核迁移至细胞质,其细胞定位的改变表明METTL3可能参与调控FMDV基因组的转录过程.本研究中成功构建了 METTL3基因敲除和过表达的BHK-21细胞系,首次发现该分子正向调控FMDV的复制.上述实验结果为进一步开展METTL3在宿主细胞中调控FMDV复制的机制研究奠定了可靠的数据基础.
Effect of METTL3 expression level on foot-and-mouth disease virus replication
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.

methyltransferase-like 3N6-methyladenosine modificationfoot-and-mouth disease virus

张继燕、李伟伟、范许许、曹雪静、朱兆宇、裴丹诗、王一卓、何路、李熙忠、任青峰、张伟、王川、郑海学

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甘肃农业大学动物医学院,甘肃兰州 730070

中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室国家口蹄疫参考实验室,甘肃兰州 730000

河南农业大学 动物医学院,河南 郑州 450046

甘肃农业大学生命科学技术学院,甘肃兰州 730070

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METTL3 m6A修饰 口蹄疫病毒

国家自然科学基金青年项目国家自然科学基金青年项目国家重点研发计划项目国家重点研发计划项目甘肃省青年科技基金计划项目中国博士后科学基金项目国家生猪产业技术体系项目国家生猪产业技术体系项目国家生猪技术创新中心先导科技项目中国农业科学院创新计划项目中国农业科学院创新计划项目"十四五"广东省农业科技创新十大主攻方向"揭榜挂帅"项目兰州兽医研究所基本科研业务费项目

32202781323028572021YFD18013002021YFD180030022JR5RA0342023M743830CARS-35CARS-39-13NCTIP-XD/C03CAAS-CSLPDCP-2023002CAAS-ASTIP-2023-LVRI2023SDZG021610312021009

2024

10.16656/j.issn.1673-4696.2024.0173

中国兽医科学
中国农业科学院兰州兽医研究所

中国兽医科学

CSTPCD北大核心
影响因子：0.524
ISSN：1673-4696
年,卷(期)：2024.54(8)
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