A cell-free platform to measure coronavirus membrane fusion
Summary: Here we present a protocol to measure coronavirus-mediated membrane fusion, an essential event in coronavirus cell entry. The approach uses nanoluciferase (Nluc) “HiBiT”-tagged corona virus-like particles (VLPs) and Nluc “LgBiT”-containing extracellular vesicles (EVs) as proxies for virus a...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723001478 |
| _version_ | 1827981824950796288 |
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| author | Thomas Kicmal Enya Qing Grant M. Hawkins Alexandria Wilcox Tom Gallagher |
| author_facet | Thomas Kicmal Enya Qing Grant M. Hawkins Alexandria Wilcox Tom Gallagher |
| author_sort | Thomas Kicmal |
| collection | DOAJ |
| description | Summary: Here we present a protocol to measure coronavirus-mediated membrane fusion, an essential event in coronavirus cell entry. The approach uses nanoluciferase (Nluc) “HiBiT”-tagged corona virus-like particles (VLPs) and Nluc “LgBiT”-containing extracellular vesicles (EVs) as proxies for virus and cell, respectively. VLP-EV membrane fusion allows HiBiT and LgBiT to combine into measurable Nluc, which signifies virus fusion with target cell membranes. We highlight assay utility with methods to assess coronavirus-mediated fusion and its inhibition by antibodies and antiviral agents.For complete details on the use and execution of this protocol, please refer to Qing et al. (2021),1 Qing et al. (2022),2 and Marcink et al. (2022).3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-09T22:15:12Z |
| format | Article |
| id | doaj.art-87d263e8dcbd427399a3a7ae544a91ac |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-09T22:15:12Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-87d263e8dcbd427399a3a7ae544a91ac2023-03-23T04:37:01ZengElsevierSTAR Protocols2666-16672023-06-0142102189A cell-free platform to measure coronavirus membrane fusionThomas Kicmal0Enya Qing1Grant M. Hawkins2Alexandria Wilcox3Tom Gallagher4Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA; Corresponding authorDepartment of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USADepartment of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USADepartment of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USADepartment of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA; Corresponding authorSummary: Here we present a protocol to measure coronavirus-mediated membrane fusion, an essential event in coronavirus cell entry. The approach uses nanoluciferase (Nluc) “HiBiT”-tagged corona virus-like particles (VLPs) and Nluc “LgBiT”-containing extracellular vesicles (EVs) as proxies for virus and cell, respectively. VLP-EV membrane fusion allows HiBiT and LgBiT to combine into measurable Nluc, which signifies virus fusion with target cell membranes. We highlight assay utility with methods to assess coronavirus-mediated fusion and its inhibition by antibodies and antiviral agents.For complete details on the use and execution of this protocol, please refer to Qing et al. (2021),1 Qing et al. (2022),2 and Marcink et al. (2022).3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723001478Cell BiologyMicrobiologyMolecular Biology |
| spellingShingle | Thomas Kicmal Enya Qing Grant M. Hawkins Alexandria Wilcox Tom Gallagher A cell-free platform to measure coronavirus membrane fusion STAR Protocols Cell Biology Microbiology Molecular Biology |
| title | A cell-free platform to measure coronavirus membrane fusion |
| title_full | A cell-free platform to measure coronavirus membrane fusion |
| title_fullStr | A cell-free platform to measure coronavirus membrane fusion |
| title_full_unstemmed | A cell-free platform to measure coronavirus membrane fusion |
| title_short | A cell-free platform to measure coronavirus membrane fusion |
| title_sort | cell free platform to measure coronavirus membrane fusion |
| topic | Cell Biology Microbiology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723001478 |
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