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Pseudotyped VSV-SARS-CoV-2 Spike

The glycoprotein (G protein) of VSV is responsible for the attachment and entry of VSV into a susceptible host cell and is therefore essential for virus infectivity. To identify and dissect the signals required for the replication and assembly of VSV, a "reverse genetics" system has been developed in which the G protein of VSV has been deleted (rVSV-ΔG). rVSV-ΔG has been used to produce VSV pseudovirus containing the envelope glycoproteins of heterologous viruses including viruses that require high-level containment (e.g., ebola viruses, SARS coronavirus, SARS-CoV-2, MERS coronavirus, etc.).

Studying how coronavirus uses its S protein to enter host cells is important for developing prophylactic and therapeutic regimes. However, the use of live virus is technically challenging in cases of dangerous strains such as SARS-CoV-2, as it requires biosafety level 3 (BSL-3) labs that are not available to most researchers. An alternative approach is to use recombinant vesicular stomatitis virus (VSV) pseudotyped with the coronavirus S protein in cell entry assays.

Product Description

Pseudotyped VSV-SARS-CoV-2 S-ΔG encodes the antigenomic-sense (or positive-sense) RNA of a replicaton-restricted recombinant vesicular stomatitis virus (rVSV) in which the glycoprotein (G) gene has been replaced with SARS-CoV-2 spike protein. The pseudotype VSV particles encode reporter gene (mCherry/GFP/Luciferase) together with the VSV nucleocapsid (N), phosphoprotein (P), and large polymerase subunit (L) in their pVSV-ΔG vector. When the VSV pseudovirus infect the target cells, reporter gene expression is proportional to the number of cells that were infected.

Fig. 1 Pseudotyped rVSV-ΔG-spike design and generation strategy

Experimental Validation

Infectivity determined in ACE2/293T using serial dilutions of Pseudotyped VSV-SARS-CoV-2 S-ΔG in a 96-well plate:

Pseudovirus Input (uL)

Product List

Cat# Product Name Reporter Gene Envelope Glycoproteins
COV-PSV10 Pseudotyped VSV-SARS-CoV-2 S-ΔG-mCherry mCherry SARS-CoV-2 Spike (Wuhan-Hu-1)
COV-PSV11 Pseudotyped VSV-SARS-CoV-2 S-ΔG-EGFP GFP SARS-CoV-2 Spike (Wuhan-Hu-1)
COV-PSV12 Pseudotyped VSV-SARS-CoV-2 S-ΔG-Luciferase Luciferase SARS-CoV-2 Spike (Wuhan-Hu-1)

Pseudotyped Luciferase/GFP VSV is designed as a control for CD's Pseudotyped Luciferase/GFP rSARS-CoV-2 Spike to test for non-specific factors that affect virus infectivity. The Pseudovirus display the VSV envelope glycoprotein (VSV-G) pseudotyped on replication-incompetent virus particles that contain a heterologous lentiviral (HIV) core. The VSV-G protein confers the pseudovirus with a high level of single cycle infectivity due to its broad tropism.

Cat# Product Name Reporter Gene Envelope Glycoproteins
VSV-PS01 Pseudotyped Luciferase VSV Luciferase VSV-G
VSV-PS02 Pseudotyped GFP VSV GFP VSV-G

SARS-CoV-2 Variant rVSV-ΔG Pseudovirus

Cat. No. SARS-CoV-2 / Variant WHO Mutations
COV-PSV13 D614G ——— D614G
COV-PSV14 Indian variant B.1.617.1 Kappa G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H
COV-PSV15 Indian variant B.1.617.2 Delta T19R, G142D, del156/157, R158G, L452R, T478K, D614G, P681R, D950N
COV-PSV16 C.37 Lambda G75V, T76I, R246N, del247-253, L452Q, F490S, D614G, T859N
COV-PSV17 Brazilian variant P.1, 20J/501Y.V3 Gamma ΔH69/V70, ΔY144, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H
COV-PSV18 South African variant Δ3 B.1.351, 20H/501Y.V2 Beta L18F, D80A, D215G, ΔL242/A243/L244, R246I, K417N, N501Y, E484K, D614G, A701V
COV-PSV19 UK variant B.1.1.7, 20I/501Y.V1 Alpha ΔH69/V70, ΔY144, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H

Human Covirovirus rVSV-ΔG Pseudovirus

Cat. No. Virus Envelope Glycoproteins
COV-PSV20 SARS-CoV S spike protein [Gene ID: 1489668]
COV-PSV21 MERS-CoV S spike protein [Gene ID: 14254594]
COV-PSV22 Human coronavirus OC43 S spike surface glycoprotein [Gene ID: 39105218]
COV-PSV23 Human coronavirus HKU1 S spike glycoprotein [Gene ID: 3200426]

Gene Reporters Available

Gene reporters for pseudotyping Methodology of detection Advantages Disadvantages
Pseudotyped ΔG-GFP (G*ΔG-GFP) rVSV Fluorescence Live-cell imaging;
Cytotoxicity evaluation;
Cells usable for RNA/protein work;
Flourescence microscope
Pseudotyped ΔG-mCherry (G*ΔG-mCherry ) rVSV Fluorescence Live-cell imaging;
Cytotoxicity evaluation;
Cells usable for RNA/protein work;
Flourescence microscope
Pseudotyped ΔG-Luciferase (G*ΔG-Luciferase) rVSV Luminescence assay Low cost for kits and reagents;
Plate reader
Background for pseudoviral infection

Click here to view our Human ACE2 Stable Cell Line for coronavirus research

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