The Spike (SARS-CoV-2) pseudotyped lentivirus was produced with SARS-CoV-2 Spike protein as the envelope glycoprotein instead of vesicular stomatitis G (VSV-G) envelope glycoprotein. These pseudoviruses also contain Luciferase gene driven by CMV promoter and GFPT2A-puro driven by EF1α promoter, therefore, the spike-mediated cell entry can be conveniently quantified by luciferase activity and visualized under a fluorescent microscope. The Spike (SARS-CoV-2) pseudotyped lentivirus can be used for studying basic mechanisms and drug development including measuring the activity of neutralizing antibody against SARS-CoV-2, and screening anti-CoV2 drugs in a Biosafety Level 2 facility

Background

The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Infection of SARS-CoV-2 to human cells is initiated by the binding of its Spike protein to ACE2 (angiotensin converting enzyme 2), a homodimer receptor, on the surface of some human cells, particularly those in the respiratory tract. The Spike protein promotes attachment of ACE2 to the subunit S1 and its RBD (Receptor Binding Domain) region, facilitates fusion of the viral and cellular membrane releasing the virus core into the cell. Therefore, drugs targeting the interaction between the Spike protein of SARSCoV-2 and ACE2 may offer protection against the viral infection. Due to the highly transmissible and pathogenic nature of SARS-CoV-2, handling of live virus requires a biosafety level 3 (BSL3) containment. In order to extend this capability to BSL-2, a high titer pseudotype virus is needed to meet such requirement that effectively replaces the need for the live SARS-CoV-2.

Literature:

1. Crawford, K.H.D., et al., Viruses, 2020. 12(5).