Spike (BA.2.86, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentivirus (eGFP Reporter)

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78982

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Description

The Spike (BA.2.86 Omicron Variant) (SARS-CoV-2) Pseudotyped Lentivirus (eGFP Reporter) are replication incompetent, HIV-based lentiviral particles. They were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BA.2.86 mutations; see below for details) as the envelope glycoprotein, instead of the commonly used VSV-G. These pseudovirions also contain the eGFP reporter driven by a CMV promoter (Figure 1), allowing to measure spike-mediated cell entry using the eGFP fluorescence signal.

The pseudovirions have been validated in a cellular assay with ACE2-HEK293 Recombinant Cell Line (#79951), a cell line that overexpresses ACE2 at high levels, as target cell line.

Figure 1. Schematic of the lenti-vector used to introduce the eGFP Reporter in the Spike (BA.2.86, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentivirus (eGFP Reporter).

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Fluorescence microscopy of ACE2-HEK293 cells transduced with Spike (BA.2.86, Omicron Variant) Pseudotyped Lentivirus (eGFP Reporter).

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, 78982-1 (100 µl size) provides sufficient pseudovirions to create a signal-to-noise ratio that allows to perform 100 reactions, and 78982-2 (500 µl x 2) for 1000 reactions. The amount of virus added to the cells can 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
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939
96-well white clear-bottom tissue culture plate	Corning #3610

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

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.

Spike Mutations in BA.2.86 Omicron Variant:
T19I, LPP24-26del, A27S, S50L, HV69-70del, V127F, G142D, Y144del, F157S, R158G, N211del, L212I, V213G, L216F, H245N, A264D, I332V, G339H, K356T, S371F, S373P, S375F, T376A, R403K, D405N, R408S, K417N, N440K, V445H, G446S, N450D, L452W, N460K, S477N, T478K, N481K, V483del, E484K, F486P, Q498R, N501Y, Y505H, E554K, A570V, D614G, P621S, H655Y, N679K, P681R, N764K, D796Y, S739F, Q954H, N969K, P1143L

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Applications

Screen or titrate neutralizing antibodies against the SARS-CoV-2 Spike Omicron BA.2.86 variant in ACE2-expressing cells.

Shipping Temperature

-80°C

Notes

Biosafety
None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Coronavirus

Regulatory Status

Research Use Only

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Spike (BA.2.86, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentivirus (eGFP Reporter)

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