Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies

Surbhi Dahiya; Sudhakar Singh; Sharvan Sehrawat

doi:10.1016/j.xpro.2022.101977

Protocol for investigating the biogenesis of SARS-CoV-2 S pseudoviruses in HEK293T cells transduced to express the virus-specific intrabodies

STAR Protoc. 2023 Mar 17;4(1):101977. doi: 10.1016/j.xpro.2022.101977. Epub 2022 Dec 15.

Authors

Surbhi Dahiya¹, Sudhakar Singh¹, Sharvan Sehrawat²

Affiliations

¹ Department of Biological Sciences, Indian Institute of Science Education and Research Mohali, Sector 81, SAS Nagar Knowledge City PO Manauli, Mohali, Punjab 140306, India.
² Department of Biological Sciences, Indian Institute of Science Education and Research Mohali, Sector 81, SAS Nagar Knowledge City PO Manauli, Mohali, Punjab 140306, India. Electronic address: sharvan@iisermohali.ac.in.

Abstract

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.

Keywords: Cell Biology; Immunology; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19*
Genetic Vectors / genetics
HEK293 Cells
Humans
Lentivirus / genetics
SARS-CoV-2* / genetics