Live cell assay to quantify the activity of SARS-CoV-2 main protease, Mpro

Fluorescence-based method to effectively screen for protease inhibitors
Technology No. 2021-102
IP Status: US Patent Pending


  • Screening, characterization, and development of protease inhibitors against SARS-CoV-2 and related coronaviruses

Key Benefits & Differentiators

  • Robust quantification system: live cell, fluorescence-based gain-of-function assay for inhibitor screening
  • High specificity and sensitivity: exquisite specificity and high signal-to-noise ratio

Technology Summary

Despite the ongoing COVID-19 pandemic, there are still no effective drugs to treat disease caused by SARS-CoV-2. SARS-CoV-2 viral proteases (Mpro or PLpro) and the RNA dependent RNA polymerase (RdRp) are ideal targets for drug discovery. The main protease, Mpro, is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis and is therefore an attractive drug target. A wide range of biochemical assays are available for measuring SARS-CoV-2 protease activity, but specific and sensitive cellular assays are less developed. There are currently no sensitive and specific live cell assays available for quantifying SARS-CoV-2 Mpro inhibition, which hinders development of drugs to target this enzyme.

Researchers at the University of Minnesota have developed a quantitative reporter for Mpro function in living cells (Src-Mpro-Tat-eGFP), in which protease inhibition by genetic or chemical methods results in strong eGFP fluorescence (Fig. 1). This robust gain-of-function system readily distinguishes between inhibitor potencies and can be scaled-up to high-throughput platforms for drug development. Essentially, the better the chemical inhibitor/drug, the stronger the fluorescent readout of the assay which expedites any high-throughput readout.

Phase of Development

TRL: 4-6
Proof of concept with commercial inhibitors and tested in 293, 293T, HeLa and U2OS cell lines.

Desired Partnerships

This technology is now available for:
  • License
  • Sponsored research
  • Co-development

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  • swap_vertical_circlelibrary_booksReferences (1)
    1. Moghadasi, S.A., Becker, J.T., Belica, C., Wick, C., Brown, W.L. and Harris, R.S. (2020), Gain-of-function assay for SARS-CoV-2 Mpro inhibition in living cells, bioRxiv
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