Spike (B.1.1.7, Alpha Variant) (SARS-CoV-2) Pseudotyped Lentivirus (eGFP Reporter)
The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein and ACE2 may offer protection against the viral infection. The United Kingdom (UK) identified a variant called B.1.1.7 with a large number of mutations in the fall of 2020. This variant spreads more easily and quickly than other variants.
The Spike (B.1.1.7 Variant) (SARS-CoV-2) Pseudotyped Lentivirus were produced with SARS-CoV-2 B.1.1.7 variant Spike (Genbank Accession #QHD43416.1 with B.1.1.7 variant mutations; see below for details) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions contain the enhanced green fluorescent protein (eGFP) gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be conveniently determined via eGFP fluorescence. The Spike (B.1.1.7 Variant) (SARS-CoV-2) pseudotyped lentivirus can be used to measure the activity of neutralizing antibody against SARS-CoV-2 B.1.1.7 variant in a Biosafety Level 2 facility.
Spike interacts with host cells by binding to membrane receptor ACE2 (angiotensin converting enzyme 2). Based on experiments performed by scientists at BPS Bioscience, we know that the wild-type SARS-CoV-2 spike pseudotyped lentiviruses transduce the following cells with great efficiency: ACE2-HEK293 cells (BPS Bioscience, #79951), ACE2-CHO cells (BPS Bioscience, #79959), ACE2-HeLa cells (BPS Bioscience, #79958). They also efficiently transduce TMPRSS2-Vero E6 cells (BPS Bioscience, #78081), which express high endogenous levels of ACE2 and were stably transfected with human serine protease TMPRSS2 required for the priming of Spike and fusion of the virion with the plasma membrane. By contrast, it has been shown by others that SARS-CoV-2 spike pseudotyped lentiviruses do not transduce parental Calu3 and Vero E6 cells very well [Neerukonda et al. 2021, PlosOne PMID: 33690649; Tandon et al. 2020, Scientific Reports PMID: 33154514; Condor Capcha et al. 2021, Front. Cardiovasc. Med. PMID: 33521067; Pisil et al. 2021, Pathogens PMID: 33540924].
SARS-CoV-2 variant pseudoviruses have been validated using ACE2-HEK293 cells but have not been tested in other cells.
Spike Mutations in B.1.1.7 variant
Deletions of H69, V70, and Y144
N501Y
A570D
D614G
P681H
T716I
S982A
D1118H
Figure 1. Schematic of the eGFP Reporter in Spike (B.1.1.7 Variant) (SARS-CoV-2) Pseudotyped Lentivirus
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| Name | Ordering Information |
| Thaw Medium 1 | BPS Bioscience, #60187 |
| ACE2- HEK293 Recombinant Cell Line | BPS Bioscience, #79951 |
| 96-well white clear-bottom assay plate | Corning, #3610 |
The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS.
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