Spike (BA.4/5, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentivirus (Luc Reporter)

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78651
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Description

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 of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants have been divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. Among them, BA.4 and BA.5 have identical mutations on their spike protein. The spike protein of BA.4 and BA.5 are referred as BA.4/5 in this datasheet.

The Spike (BA.4/5, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BA.4/5 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BA.4/5, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BA.4/5 variant in a Biosafety Level 2 facility.

As shown in Figures 2 and 3, the Spike Omicron BA.4/5 pseudovirus has been validated for use with target cells ACE2-HEK293 (which overexpress ACE2; BPS Bioscience #79951).

Spike Mutations in BA.4/5, Omicron Variant:

Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K

Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion

Product Info
Storage and Usage
Citations
Supplied As
The titer will vary with each lot; the exact value is provided with each shipment. Based on experiments performed by scientists at BPS Bioscience, 78651-1 (100 µl) provides sufficient signal-to-noise ratio to perform 100 reactions, and 78651-2 (500 µl x 2) for 1000 reactions. The amount of virus added to the cells can even be titrated further down according to the user’s need.
Materials Required But Not Supplied
Name Ordering Information
Thaw Medium 1  BPS Bioscience #60187
ACE2-HEK293 Recombinant Cell Line BPS Bioscience #79951
Spike Neutralizing Antibody (Clone G10xA1) (SARS-CoV-2) BPS Bioscience #101326
ONE-Step™ luciferase assay system BPS Bioscience #60690
96-well tissue culture treated, white clear-bottom assay plate Corning, #3610
Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.