A genome-wide CRISPR/Cas9 knockout screen identifies TMEM239 as an important host factor in facilitating African swine fever virus entry into early endosomes

文献类型： 外文期刊

第一作者： Shen, Dongdong

作者： Shen, Dongdong;Liu, Renqiang;Sheng, Xiangpeng;Zhu, Yuanmao;Sun, Encheng;Zhang, Jiwen;Li, Fang;Xia, Changyou;Bu, Zhigao;Zhao, Dongming;Shen, Dongdong;Ge, Junwei;Zhang, Guigen;Weng, Xiaogang;Zhang, Yuting;Liu, Yan;Mu, Yanshuang;Liu, Zhonghua;Liu, Zhiheng

作者机构：

期刊名称：PLOS PATHOGENS （ 影响因子：5.5； 五年影响因子：5.5 ）

ISSN： 1553-7366

年卷期： 2024 年 20 卷 7 期

页码：

收录情况： SCI

摘要： African swine fever (ASF) is a highly contagious, fatal disease of pigs caused by African swine fever virus (ASFV). The complexity of ASFV and our limited understanding of its interactions with the host have constrained the development of ASFV vaccines and antiviral strategies. To identify host factors required for ASFV replication, we developed a genome-wide CRISPR knockout (GeCKO) screen that contains 186,510 specific single guide RNAs (sgRNAs) targeting 20,580 pig genes and used genotype II ASFV to perform the GeCKO screen in wild boar lung (WSL) cells. We found that knockout of transmembrane protein 239 (TMEM239) significantly reduced ASFV replication. Further studies showed that TMEM239 interacted with the early endosomal marker Rab5A, and that TMEM239 deletion affected the co-localization of viral capsid p72 and Rab5A shortly after viral infection. An ex vivo study showed that ASFV replication was significantly reduced in TMEM239-/- peripheral blood mononuclear cells (PBMCs) from TMEM239 knockout piglets. Our study identifies a novel host factor required for ASFV replication by facilitating ASFV entry into early endosomes and provides insights for the development of ASF-resistant breeding. ASFV is a major threat to pig industry worldwide and relies on host factors to complete its replication cycle. We developed a genome-wide CRISPR knockout (GeCKO) screen and identified TMEM239, a member of the transmembrane (TMEM) protein family, as a potential host factor required for ASFV replication. TMEM239 interacts with the early endosome marker Rab5A and TMEM239 knockout impedes ASFV entry into early endosomes. We further generated TMEM239 knockout pigs and ASFV replication was significantly reduced in TMEM239-/- peripheral blood mononuclear cells (PBMCs). Our findings will contribute to the development of strategies for pig breeding against ASF.

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