Genome-wide CRISPR screen reveals key role of sialic acids in PEDV and porcine coronavirus infections

作者：Guanghao Guo, Mengjia Zhang, Zhuojia Xu, Peng Xi, Hongmei Zhu, Anouk Evers, Robert Jan Lebbink, Yifei Lang, Qigai He, Yao‐Wei Huang, Tiehai Li, Berend‐Jan Bosch, Wentao Li · 发表于：mBio · 年份：2025 · DOI：10.1128/mbio.01628-25 · 被引用次数：8 · 研究领域：Animal Virus Infections Studies、Viral gastroenteritis research and epidemiology、SARS-CoV-2 and COVID-19 Research

Porcine epidemic diarrhea virus (PEDV) is a globally distributed alphacoronavirus with economic importance that can cause severe watery diarrhea and even death in piglets. To identify host factors essential for PEDV infection, we performed a genome-wide CRISPR/Cas9 screen in human hepatocellular carcinoma cells (Huh7) using the highly virulent PEDV GIIb strain GDU. Several genes involved in the sialic acid and heparan sulfate biosynthesis pathway and cholesterol metabolism were highly enriched following PEDV selection. We validated that the host factor ST3 beta-galactoside alpha-2,3-sialyltransferase 4 (ST3GAL4), which catalyzes the transfer of sialic acid to sugar chains via α2,3-linked linkages, is important for PEDV infection. To systematically investigate the role of sialic acid in PEDV infection, we knocked out genes related to sialic acid synthesis. This led to a reduced abundance of sialic acid on the cell surface, which in turn inhibited PEDV adsorption and internalization. Furthermore, we found that both α2,3-linked and α2,6-linked sialic acids can serve as cellular attachment factors for PEDV. We conducted a glycan microarray screen to determine which sialoglycans are preferred by the PEDV spike protein. The results revealed that PEDV favors binding to α2,3-sialoglycans. Additionally, we found that not only current circulating PEDV strains but also other porcine coronaviruses rely on sialic acid for efficient infection. Collectively, our findings provide insights in...