J Biol Chem . SARS-CoV-2 viral budding and entry can be modeled using BSL-2 level virus-like particles

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J Biol Chem


. 2020 Nov 19;jbc.RA120.016148.
doi: 10.1074/jbc.RA120.016148. Online ahead of print.
SARS-CoV-2 viral budding and entry can be modeled using BSL-2 level virus-like particles


Caroline B Plescia ¹ , Emily A David ¹ , Dhabaleswar Patra ¹ , Ranjan Sengupta ² , Souad Amiar ³ , Yuan Su ¹ , Robert V Stahelin ¹



Affiliations

• PMID: 33214224
• DOI: 10.1074/jbc.RA120.016148

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first discovered in December 2019 in Wuhan, China and expeditiously spread across the globe causing a global pandemic. Research on SARS-CoV-2, as well as the closely related SARS-CoV-1 and MERS coronaviruses is restricted to BSL-3 facilities. Such BSL-3 classification make SARS-CoV-2 research inaccessible to the majority of functioning research laboratories in the US; this becomes problematic when the collective scientific effort needs to be focused on such in the face of a pandemic. However, a minimal system capable of recapitulating different steps of the viral life cycle without using the virus' genetic material could increase accessibility. In this work, we assessed the four structural proteins from SARS-CoV-2 for their ability to form virus-like particles (VLPs) from human cells to form a competent system for BSL-2 studies of SARS-CoV-2. Herein, we provide methods and resources of producing, purifying, fluorescently and APEX2-labeling of SARS-CoV-2 VLPs for the evaluation of mechanisms of viral budding and entry as well as assessment of drug inhibitors under BSL-2 conditions. These systems should be useful to those looking to circumvent BSL-3 work with SARS-CoV-2 yet study the mechanisms by which SARS-CoV-2 enters and exits human cells.

Keywords: Golgi; SARS-CoV-2; coronavirus; electron microscopy (EM); endoplasmic reticulum (ER); membrane bilayer; virology; virus; virus assembly; virus budding; virus entry; virus-like particle.