Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies
Summary: The protocol is designed to investigate the influence of an anti-cleavage site intrabody in modulating the output of LV(CoV-2 S), a lentivirus-based pseudovirus expressing CoV-2 S protein using HEK293T cells. We clone the single-domain antibody sequence into a lentiviral vector (pLenti-GFP)...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722008577 |
| _version_ | 1797961034117414912 |
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| author | Surbhi Dahiya Sudhakar Singh Sharvan Sehrawat |
| author_facet | Surbhi Dahiya Sudhakar Singh Sharvan Sehrawat |
| author_sort | Surbhi Dahiya |
| collection | DOAJ |
| description | Summary: The protocol is designed to investigate the influence of an anti-cleavage site intrabody in modulating the output of LV(CoV-2 S), a lentivirus-based pseudovirus expressing CoV-2 S protein using HEK293T cells. We clone the single-domain antibody sequence into a lentiviral vector (pLenti-GFP) for intracellular expression and assess not only the viral biogenesis but also the fate of the CoV-2 S protein in such cells.For complete details on the use and execution of this protocol, please refer to Singh et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T00:53:03Z |
| format | Article |
| id | doaj.art-33264c4ead954ddeb267055934522eee |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T00:53:03Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-33264c4ead954ddeb267055934522eee2023-01-05T07:03:39ZengElsevierSTAR Protocols2666-16672023-03-0141101977Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodiesSurbhi Dahiya0Sudhakar Singh1Sharvan Sehrawat2Department of Biological Sciences, Indian Institute of Science Education and Research Mohali, Sector 81, SAS Nagar Knowledge City PO Manauli, Mohali, Punjab 140306, IndiaDepartment of Biological Sciences, Indian Institute of Science Education and Research Mohali, Sector 81, SAS Nagar Knowledge City PO Manauli, Mohali, Punjab 140306, IndiaDepartment of Biological Sciences, Indian Institute of Science Education and Research Mohali, Sector 81, SAS Nagar Knowledge City PO Manauli, Mohali, Punjab 140306, India; Corresponding authorSummary: The protocol is designed to investigate the influence of an anti-cleavage site intrabody in modulating the output of LV(CoV-2 S), a lentivirus-based pseudovirus expressing CoV-2 S protein using HEK293T cells. We clone the single-domain antibody sequence into a lentiviral vector (pLenti-GFP) for intracellular expression and assess not only the viral biogenesis but also the fate of the CoV-2 S protein in such cells.For complete details on the use and execution of this protocol, please refer to Singh et al. (2022).1 : 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/S2666166722008577Cell BiologyImmunologyMolecular Biology |
| spellingShingle | Surbhi Dahiya Sudhakar Singh Sharvan Sehrawat Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies STAR Protocols Cell Biology Immunology Molecular Biology |
| title | Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies |
| title_full | Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies |
| title_fullStr | Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies |
| title_full_unstemmed | Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies |
| title_short | Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies |
| title_sort | protocol for investigating the biogenesis of sars cov 2 s pseudoviruses in hek293t cells transduced to express the virus specific intrabodies |
| topic | Cell Biology Immunology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722008577 |
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