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Nanoluciferase complementation-based bioreporter reveals the importance of N-linked glycosylation of SARS-CoV-2 S for viral entry

Mol Ther. 2021-02; 
Taha Azad, Ragunath Singaravelu, Zaid Taha, Taylor R Jamieson, Stephen Boulton, Mathieu J F Crupi, Nikolas T Martin, Emily E F Brown, Joanna Poutou, Mina Ghahremani, Adrian Pelin, Kazem Nouri, Reza Rezaei, Christopher Boyd Marshall, Masahiro Enomoto, Rozanne Arulanandam, Nouf Alluqmani, Reuben Samson, Anne-Claude Gingras, D William Cameron, Peter A Greer, Carolina S Ilkow, Jean-Simon Diallo, John C Bell
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PCR and Cloning Inserts outlined in Table S1 were ordered from GenScript Get A Quote

摘要

The ongoing COVID-19 pandemic has highlighted the immediate need for the development of antiviral therapeutics targeting different stages of the SARS-CoV-2 life cycle. We developed a bioluminescence-based bioreporter to interrogate the interaction between the SARS-CoV-2 viral spike (S) protein and its host entry receptor, angiotensin-converting enzyme 2 (ACE2). The bioreporter assay is based on a nanoluciferase complementation reporter, composed of two subunits, large BiT and small BiT, fused to the S receptor-binding domain (RBD) of the SARS-CoV-2 S protein and ACE2 ectodomain, respectively. Using this bioreporter, we uncovered critical host and viral determinants of the interaction, including a role for glyc... More

关键词

2019-nCoV, COVID-19, SARS-CoV-2, bioluminescence, bioreporter, coronavirus, high-throughput screening, viral entry