CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex
Abstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, TMPRSS2 represents a key factor for SARS-CoV-2 infection, as TMPRSS2 mediates cleavage of viral spike protein, enabling direct fusion of the viral env...
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Nature Portfolio
2025-04-01
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| Online Access: | https://doi.org/10.1038/s41598-025-95644-0 |
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| author | Ildar Gabaev Alexandra Rowland Emilija Jovanovic Christian M. Gawden-Bone Thomas W. M. Crozier Ana Teixeira-Silva Edward J. D. Greenwood Pehuén Pereyra Gerber Niek Wit James A. Nathan Nicholas J. Matheson Paul J. Lehner |
| author_facet | Ildar Gabaev Alexandra Rowland Emilija Jovanovic Christian M. Gawden-Bone Thomas W. M. Crozier Ana Teixeira-Silva Edward J. D. Greenwood Pehuén Pereyra Gerber Niek Wit James A. Nathan Nicholas J. Matheson Paul J. Lehner |
| author_sort | Ildar Gabaev |
| collection | DOAJ |
| description | Abstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, TMPRSS2 represents a key factor for SARS-CoV-2 infection, as TMPRSS2 mediates cleavage of viral spike protein, enabling direct fusion of the viral envelope with the host cell membrane. Since the start of the COVID-19 pandemic, TMPRSS2 has gained attention as a therapeutic target for protease inhibitors which would inhibit SARS-CoV-2 infection, but little is known about TMPRSS2 regulation, particularly in cell types physiologically relevant for SARS-CoV-2 infection. Here, we performed an unbiased genome-wide CRISPR-Cas9 library screen, together with a library targeted at epigenetic modifiers and transcriptional regulators, to identify cellular factors that modulate cell surface expression of TMPRSS2 in human colon epithelial cells. We find that endogenous TMPRSS2 is regulated by the Elongin BC-VHL complex and HIF transcription factors. Depletion of Elongin B or treatment of cells with PHD inhibitors resulted in downregulation of TMPRSS2 and inhibition of SARS-CoV-2 infection. We show that TMPRSS2 is still utilised by SARS-CoV-2 Omicron variants for entry into colonic epithelial cells. Our study enhances our understanding of the regulation of endogenous surface TMPRSS2 in cells physiologically relevant to SARS-CoV-2 infection. |
| format | Article |
| id | doaj-art-934183e0b64f4e1e9e0e0db771464a74 |
| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-934183e0b64f4e1e9e0e0db771464a742025-08-20T03:06:57ZengNature PortfolioScientific Reports2045-23222025-04-0115111710.1038/s41598-025-95644-0CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complexIldar Gabaev0Alexandra Rowland1Emilija Jovanovic2Christian M. Gawden-Bone3Thomas W. M. Crozier4Ana Teixeira-Silva5Edward J. D. Greenwood6Pehuén Pereyra Gerber7Niek Wit8James A. Nathan9Nicholas J. Matheson10Paul J. Lehner11Department of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeWellcome-MRC Cambridge Stem Cell Institute, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeDepartment of Medicine, University of CambridgeAbstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, TMPRSS2 represents a key factor for SARS-CoV-2 infection, as TMPRSS2 mediates cleavage of viral spike protein, enabling direct fusion of the viral envelope with the host cell membrane. Since the start of the COVID-19 pandemic, TMPRSS2 has gained attention as a therapeutic target for protease inhibitors which would inhibit SARS-CoV-2 infection, but little is known about TMPRSS2 regulation, particularly in cell types physiologically relevant for SARS-CoV-2 infection. Here, we performed an unbiased genome-wide CRISPR-Cas9 library screen, together with a library targeted at epigenetic modifiers and transcriptional regulators, to identify cellular factors that modulate cell surface expression of TMPRSS2 in human colon epithelial cells. We find that endogenous TMPRSS2 is regulated by the Elongin BC-VHL complex and HIF transcription factors. Depletion of Elongin B or treatment of cells with PHD inhibitors resulted in downregulation of TMPRSS2 and inhibition of SARS-CoV-2 infection. We show that TMPRSS2 is still utilised by SARS-CoV-2 Omicron variants for entry into colonic epithelial cells. Our study enhances our understanding of the regulation of endogenous surface TMPRSS2 in cells physiologically relevant to SARS-CoV-2 infection.https://doi.org/10.1038/s41598-025-95644-0Transmembrane serine proteasesCoronavirus entry factorsCRISPR-Cas9 screenColon epithelial cellsHypoxic regulation of surface proteins |
| spellingShingle | Ildar Gabaev Alexandra Rowland Emilija Jovanovic Christian M. Gawden-Bone Thomas W. M. Crozier Ana Teixeira-Silva Edward J. D. Greenwood Pehuén Pereyra Gerber Niek Wit James A. Nathan Nicholas J. Matheson Paul J. Lehner CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex Scientific Reports Transmembrane serine proteases Coronavirus entry factors CRISPR-Cas9 screen Colon epithelial cells Hypoxic regulation of surface proteins |
| title | CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex |
| title_full | CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex |
| title_fullStr | CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex |
| title_full_unstemmed | CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex |
| title_short | CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex |
| title_sort | crispr cas9 genetic screens reveal regulation of tmprss2 by the elongin bc vhl complex |
| topic | Transmembrane serine proteases Coronavirus entry factors CRISPR-Cas9 screen Colon epithelial cells Hypoxic regulation of surface proteins |
| url | https://doi.org/10.1038/s41598-025-95644-0 |
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