A cell-free platform to measure coronavirus membrane fusion

Thomas Kicmal; Enya Qing; Grant M Hawkins; Alexandria Wilcox; Tom Gallagher

doi:10.1016/j.xpro.2023.102189

A cell-free platform to measure coronavirus membrane fusion

STAR Protoc. 2023 Mar 6;4(2):102189. doi: 10.1016/j.xpro.2023.102189. Online ahead of print.

Authors

Thomas Kicmal¹, Enya Qing², Grant M Hawkins², Alexandria Wilcox², Tom Gallagher³

Affiliations

¹ Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA. Electronic address: tkicmal@luc.edu.
² Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA.
³ Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA. Electronic address: tgallag@luc.edu.

Abstract

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.

Keywords: Cell Biology; Microbiology; Molecular Biology.