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Gain-of-function assay for SARS-CoV-2 Mpro inhibition in living cells

Seyad Arad Moghadasi, View ORCID ProfileJordan T. Becker, Christopher Belica, Chloe Wick, William L. Brown, View ORCID ProfileReuben S. Harris
doi: https://doi.org/10.1101/2020.11.09.375139
Seyad Arad Moghadasi
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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Jordan T. Becker
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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Christopher Belica
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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Chloe Wick
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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William L. Brown
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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Reuben S. Harris
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455
2Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455
3Institute for Molecular Virology, University of Minnesota, Minneapolis, Minnesota, USA, 55455
4Howard Hughes Medical Institute, University of Minnesota, Minneapolis, Minnesota, USA, 55455
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  • ORCID record for Reuben S. Harris
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Abstract

The main protease, Mpro, of SARS-CoV-2 is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis. Here we demonstrate a quantitative reporter for Mpro function in living cells, in which protease inhibition by genetic or chemical methods results in strong eGFP fluorescence. This robust gain-of-function system readily distinguishes between inhibitor potencies and can be scaled-up to high-throughput platforms for drug testing.

Competing Interest Statement

RSH is a co founder, shareholder, and consultant of ApoGen Biotechnologies Inc. The other authors have declared that no competing interests exist.

Footnotes

  • ↵* E-mail: rsh{at}umn.edu

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Gain-of-function assay for SARS-CoV-2 Mpro inhibition in living cells
Seyad Arad Moghadasi, Jordan T. Becker, Christopher Belica, Chloe Wick, William L. Brown, Reuben S. Harris
bioRxiv 2020.11.09.375139; doi: https://doi.org/10.1101/2020.11.09.375139
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Gain-of-function assay for SARS-CoV-2 Mpro inhibition in living cells
Seyad Arad Moghadasi, Jordan T. Becker, Christopher Belica, Chloe Wick, William L. Brown, Reuben S. Harris
bioRxiv 2020.11.09.375139; doi: https://doi.org/10.1101/2020.11.09.375139

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