2. Academic Validation
  3. Genome-wide CRISPR screen identifies ALG5, ALG6, NF2, and FUT8 as key host proteins involved in transmissible gastroenteritis virus infection

Genome-wide CRISPR screen identifies ALG5, ALG6, NF2, and FUT8 as key host proteins involved in transmissible gastroenteritis virus infection

  • Int J Biol Macromol. 2025 Dec;333(Pt 2):148770. doi: 10.1016/j.ijbiomac.2025.148770.
Zhihua Feng 1,  Xinyan He 2,  Minhua Lin 2,  Heng Zhao 2,  Yao Chen 2,  Jianghua Chen 2,  Zhaolong Li 3,  Yangkun Shen 4,  Jianxin Chen 5,  Xiaoyu Yang 6,  Qi Chen 7
Affiliations

Affiliations

  • 1 Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, Fujian Normal University, Qishan Campus, Fuzhou, Fujian, 350117, China; College of Life Science, Fujian Normal University Qishan Campus, Fuzhou, Fujian, 350117, China; Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, Fujian, 350117, China; College of Photonic and Electronic Engineering, Fujian Normal University, Fuzhou, Fujian, 350117, China; Fuzhou Hospital of Traditional Chinese Medicine Affiliated to Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian, 350001, China.
  • 2 Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, Fujian Normal University, Qishan Campus, Fuzhou, Fujian, 350117, China; College of Life Science, Fujian Normal University Qishan Campus, Fuzhou, Fujian, 350117, China.
  • 3 Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian, 350013, China.
  • 4 Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, Fujian Normal University, Qishan Campus, Fuzhou, Fujian, 350117, China; College of Life Science, Fujian Normal University Qishan Campus, Fuzhou, Fujian, 350117, China. Electronic address: [email protected].
  • 5 Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, Fujian, 350117, China; College of Photonic and Electronic Engineering, Fujian Normal University, Fuzhou, Fujian, 350117, China. Electronic address: [email protected].
  • 6 Fuzhou Hospital of Traditional Chinese Medicine Affiliated to Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian, 350001, China. Electronic address: [email protected].
  • 7 Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, Fujian Normal University, Qishan Campus, Fuzhou, Fujian, 350117, China; College of Life Science, Fujian Normal University Qishan Campus, Fuzhou, Fujian, 350117, China; Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, Fujian, 350117, China; College of Photonic and Electronic Engineering, Fujian Normal University, Fuzhou, Fujian, 350117, China. Electronic address: [email protected].
PMID: 41205942 DOI: 10.1016/j.ijbiomac.2025.148770
Abstract

Transmissible gastroenteritis virus (TGEV) represents a significant threat to global swine production. In the absence of effective Antiviral therapies, control relies primarily on vaccination. To identify potential therapeutic targets, we performed a genome-wide CRISPR/Cas9 screen in porcine IPEC-J2 cells, which revealed asparagine-linked glycosylation 5 (ALG5), asparagine-linked glycosylation 6 (ALG6), neurofibromin 2 (NF2), and fucosyltransferase 8 (FUT8) as essential host factors for TGEV Infection. Functional characterization demonstrated that ALG5, ALG6, and NF2 knockout impaired viral adsorption and internalization through disruption of Aminopeptidase N (pAPN) transcription or N-glycosylation. Consistently, tunicamycin-mediated inhibition of N-glycosylation suppressed TGEV Infection. In contrast, FUT8 knockout specifically affects viral internalization and early replication by preventing the formation of double-membrane vesicles (DMVs) but does not affect pAPN expression. This role was independent of FUT8's fucosyltransferase activity, as the enzymatic inhibitor FDW028 had no effect. Mechanistically, we found that FUT8 interacts with the TGEV nonstructural proteins NSP3 and NSP4 to facilitate DMV biogenesis. Our findings delineate distinct mechanisms by which host factors support TGEV Infection and provide novel insights for the development of targeted Antiviral strategies.

Keywords

CRISPR screen; TGEV; host factor; infection.

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