Knockout of HDAC9 Gene Enhances Foot-and-Mouth Disease Virus Replication

文献类型： 外文期刊

第一作者： Hou, Shitong

作者： Hou, Shitong;Wang, Xiangwei;Ren, Shanhui;Meng, Xuelian;Yin, Xiangping;Zhang, Jie;Jiang, Tao;Mao, Ruoqing;Zhang, Yongguang;Sun, Yuefeng;Zhang, Jie;Tarasiuk, Kazimierz;Pejsak, Zygmunt;Mao, Ruoqing;Zhang, Yongguang;Sun, Yuefeng

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关键词： HDAC9; FMDV; CRISPR; Cas9; RNA-Seq; innate immune response

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：6.064； 五年影响因子：6.843 ）

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年卷期： 2022 年 13 卷

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收录情况： SCI

摘要： Foot-and-mouth disease virus (FMDV) is a highly contagious viral disease that mainly infects cloven-hoofed animals. Propagation of FMDV by cell culture is an important method to preserve viral biological and antigenic characteristics, which is crucial in FMD monitoring and vaccine production. However, only a few cell lines are sensitive to FMDV, and there is still a lot of room for improvement. Acetylation is an important post-translational modification, which is dynamically regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). However, the study of the relationship between FMDV and HDACs is still unclear. HDAC9 belongs to the class II of HDACs family; in this study, HDAC9 knockout (KO) BHK-21 cells were successfully established using CRISPR/cas9 technology. The results of karyotype analysis, growth curve analysis, and morphological observation showed that the HDAC9 knockout cell line was stable in growth and morphological characteristics. After infection with FMDV, the expression of viral RNA and protein, viral titers, and the copies of viral RNA in HDAC9-KO cells were significantly higher than those in NC cells. Meanwhile, RNA-seq technology was used to sequence HDAC9-KO cells and NC cells infected and uninfected with FMDV. It was found that the differentially expressed innate immune factors containing NFKBIA, SOD2, IL2RG, BCL2L1, CXCL1/2/3, and IL1RAP have significantly enriched in the Jak-STAT, NOD-like receptor, Toll-like receptor, NF-kappa B, and MAPK signaling pathway. RT-qPCR was performed to detect the expression level of differentially expressed genes and showed consistency with the RNA-seq data. These results preliminarily reveal the role of HDAC9 in host antiviral innate immune response, and the HDAC9-KO cell line could also serve as a useful tool for FMDV research.

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