Jin Wei 1 , 2 , Mia Madel Alfajaro 1 , 2 , Peter C. DeWeirdt 3 , Ruth E. Hanna 3 , William J. Lu-Culligan 4 , 5 , 6 , Wesley L. Cai 7 , Madison S. Strine 1 , 2 , Shang-Min Zhang 7 , Vincent R. Graziano 1 , 2 , Cameron O. Schmitz 1 , 2 , Jennifer S. Chen 1 , 2 , Madeleine C. Mankowski 1 , 2 , Renata B. Filler 1 , 2 , Neal G. Ravindra 8 , 9 , Victor Gasque 8 , 9 , Fernando J. de Miguel 7 , 14 , Ajinkya Patil 15 , 16 , Huacui Chen 7 , Kasopefoluwa Y. Oguntuyo 10 , Laura Abriola 11 , Yulia V. Surovtseva 11 , Robert C. Orchard 12 , Benhur Lee 10 , Brett D. Lindenbach 13 , Katerina Politi 7 , 14 , 17 , David van Dijk 8 , 9 , Cigall Kadoch 15 , 16 , Matthew D. Simon 4 , 6 , Qin Yan 7 , 14 , John G. Doench 3 , ∗ , Craig B. Wilen 1 , 2 , 14 , 18 , ∗
20 October 2020
Identification of host genes essential for SARS-CoV-2 infection may reveal novel therapeutic targets and inform our understanding of COVID-19 pathogenesis. Here, we performed genome-wide CRISPR screens in Vero-E6 cells with SARS-CoV-2, MERS-CoV, bat coronavirus HKU5 expressing the SARS-CoV-1 spike, and VSV expressing the SARS-CoV-2 spike. We identify known SARS-CoV-2 host factors including the receptor ACE2 and protease Cathepsin L. We additionally discovered pro-viral genes and pathways including HMGB1 and the SWI/SNF chromatin remodeling complex that are SARS-lineage and pan-coronavirus specific, respectively. We show HMGB1 regulates ACE2 expression and is critical for viral entry of SARS-CoV-2, SARS-CoV-1, and NL63. We also show that small molecule antagonists of identified gene products inhibited SARS-CoV-2 infection in monkey and human cells, demonstrating the conserved role of these genetic hits across species. Together this identifies potential therapeutic targets for SARS-CoV-2 and reveals SARS-lineage specific and pan-coronavirus host factors that regulate susceptibility to highly pathogenic coronaviruses.
Developed monkey CRISPR library to screen pathogenic coronaviruses in Vero-E6 cells.
Screens identified genes that are SARS-CoV-2-, MERS-CoV-, and pan-coronavirus-specific.
Therapeutic targets including SMARCA4 identified for SARS-CoV-2 infection.
HMGB1 is novel regulator of ACE2 expression and critical for viral entry.
To identify potential therapeutic targets for SARS-CoV-2 and related pathogenic coronaviruses, Wei et al. conduct genome-wide CRISPR screens in Vero-E6 cells using SARS-CoV-2, MERS-CoV or pseudoviruses presenting SARS-CoV-1 or -2 spike proteins. They identify pro-viral genes and pathways including HMGB1 and the SWI/SNF chromatin remodeling complex that are SARS-lineage and pan-coronavirus specific, respectively, and demonstrate that HMGB1 is critical for SARS-lineage viral entry because it plays a critical role in ACE2 expression.