文献类型： 外文期刊

作者： Wang, Jieru ¹ ; Liu, Hailong ² ; Yang, Yuqing ² ; Tan, Yubei ⁴ ; Sun, Limeng ⁴ ; Guo, Zishi ² ; Zeng, Xiaoyu ¹ ; Wang, Zichang ² ; Li, Sheng ² ; Yin, Lei ¹ ; Yin, Dongdong ¹ ; Shen, Xuehuai ¹ ; Dai, Yin ¹ ; Liu, Xiangdong ² ; Ruan, Jinxue ² ; Li, Xinyun ² ; Zhao, Shuhong ² ; Peng, Guiqing ⁴ ; Pan, Xiaocheng ¹ ; Wang, Chonglong ¹ ; Xie, Shengsong ² ;

作者机构： 1.Anhui Acad Agr Sci, Key Lab Pig Mol Quantitat Genet, Livestock & Poultry Epidem Dis Res Ctr Anhui Prov, Anhui Prov Key Lab Livestock & Poultry Prod Safety, Hefei 230031, Peoples R China

2.Huazhong Agr Univ, Minist Educ, Key Lab Agr Anim Genet Breeding & Reprod, Wuhan 430070, Peoples R China

3.Huazhong Agr Univ, Key Lab Swine Genet & Breeding, Minist Agr & Rural Affairs, Wuhan 430070, Peoples R China

4.Minist Agr & Rural Affairs, Key Lab Prevent & Control African Swine Fever & Ot, Wuhan 430070, Peoples R China

5.Huazhong Agr Univ, Hubei Hongshan Lab, Wuhan 430070, Peoples R China

关键词： PEDV; CRISPR screen; TRIM2; Small-molecule inhibitors

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）

ISSN： 0141-8130

年卷期： 2023 年 250 卷

页码：

收录情况： SCI

摘要： Porcine epidemic diarrhoea (PED) caused by the porcine epidemic diarrhoea virus (PEDV) is the most devastating disease in the global pig industry due to its high mortality rate in piglets. The host factors critical for PEDV replication are poorly understood. Here, we designed a pooled African green monkey genome-scale CRISPR/Cas9 knockout (VeroCKO) library containing 75,608 single guide RNAs targeting 18,993 protein-coding genes. Subsequently, we use the VeroCKO library to identify key host factors facilitating PEDV infection in Vero E6 cells. Several previously unreported genes associated with PEDV infection are highly enriched post-PEDV selection. We discovered that knocking out the tripartite motif 2 (TRIM2) and the solute carrier family 35 member A1 (SLC35A1) inhibited PEDV replication. Virtual screening and molecular docking approaches showed that chem-80,048,685 (M2) s ignificantly inhibited PEDV attachment and late replication by impeding SLC35A1. Furthermore, we found that knocking out SLC35A1 in Vero E6 cells upregulated a disintegrin and metalloprotease protein-17 (ADAM17) by splicing porcine aminopeptidase N (pAPN) and angiotensin-converting enzyme 2 (ACE2) ectodomains to reduce PEDV-infection in a CMP-Sialic Acid (CMP-SA) cell entry-independent manner. These findings provide a new perspective for a better understanding of host-pathogen interactions and new therapeutic targets for PEDV infection.