Genomewide CRISPR knockout screen identified PLAC8 as an essential factor for SADS-CoVs infection
Year: 2022
Type: article
Abstract: Zoonotic transmission of coronaviruses poses an ongoing threat to human populations. Endemic outbreaks of swine acute diarrhea syndrome coronavirus (SADS-CoV) have caused severe economic losses in the pig industry and have the potential to cause human outbreaks. Currently, there are no vaccines or specific antivirals against SADS-CoV, and our limited understanding of SADS-CoV host entry factors could hinder prompt responses to a potential human outbreak. Using a genomewide CRISPR knockout screen, we identified placenta-associated 8 protein (PLAC8) as an essential host factor for SADS-CoV infection. Knockout of PLAC8 abolished SADS-CoV infection, which was restored by complementing PLAC8 from multiple species, including human, rhesus macaques, mouse, pig, pangolin, and bat, suggesting a conserved infection pathway and susceptibility of SADS-CoV among mammals. Mechanistically, PLAC8 knockout does not affect viral entry; rather, knockout cells displayed a delay and reduction in viral subgenomic RNA expression. In a swine primary intestinal epithelial culture (IEC) infection model, differentiated cultures have high levels of PLAC8 expression and support SADS-CoV replication. In contrast, expanding IECs have low levels of PLAC8 expression and are resistant to SADS-CoV infection. PLAC8 expression patterns translate in vivo; the immunohistochemistry of swine ileal tissue revealed high levels of PLAC8 protein in neonatal compared to adult tissue, mirroring the known SADS-CoV pathogenesis in neonatal piglets. Overall, PLAC8 is an essential factor for SADS-CoV infection and may serve as a promising target for antiviral development for potential pandemic SADS-CoV.
Language: English
Subfield: Animal Science and Zoology
Domain: Life Sciences
Open Access status: hybrid
Funders: Pfizer, Chan Zuckerberg Initiative, National Institutes of Health, NIH Office of the Director, National Cancer Institute, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Environmental Health Sciences, Center for Gastrointestinal Biology and Disease, School of Medicine, University of North Carolina at Chapel Hill
Awards: G1017297203, G2328161403, G623642269, G784935900, Understanding Risk of Zoonotic Virus Emergence in EID Hotspots of Southeast Asia, G7932391634, G9000008043, G9000010008, G9000033364, G9000099626, G9000107645, G9000210059, G9000240464, G9000283304, Stanford/UNC Biomimetic U19 Research Center, Cancer Center Support Grant, UNC Cystic Fibrosis Research and Translation Core Center, G940654010
OpenAlex ID: W4225107700