VSV Pseudovirus

1. Service Introduction

Building on its advanced viral vector engineering platform, GeneRulor offers high-quality VSV pseudovirus packaging services. We employ a well-established VSV-ΔG packaging system, in which specific viral envelope glycoproteins are exogenously supplied to a VSV genome lacking its native G protein, generating pseudoviral particles with defined infectivity but no autonomous replication capacity.

The VSV pseudovirus system is an engineered viral vector based on Vesicular Stomatitis Virus (VSV). By replacing the native VSV G protein (glycoprotein) with the envelope protein of another virus of interest (e.g., SARS-CoV-2 Spike protein), a pseudovirus is produced that retains the entry characteristics of the target virus while being incapable of replication. This makes it safe for use in neutralizing antibody assays, vaccine evaluation, antiviral drug screening, and mechanistic studies of viral entry.

The workflow begins with construction of an expression plasmid encoding the target viral envelope protein (e.g., Spike protein) and a ΔG-VSV backbone plasmid (e.g., VSV-ΔG-Luciferase, carrying a reporter gene). A high-titer, high-purity working stock of VSV-ΔG/luc-G pseudovirus is then generated by large-scale amplification in HEK293T or BHK-21 cells, followed by concentration and purification. This working stock is subsequently used for packaging pseudoviruses bearing the envelope proteins of other pathogens. In brief: HEK293T or BHK-21 cells are transfected with the target envelope protein expression plasmid by lipofection, then infected with the VSV-ΔG/luc-G working stock. Supernatant is harvested 48–72 hours post-infection, clarified through a 0.45 μm filter to remove cell debris, and optionally concentrated by ultracentrifugation, yielding the final high-titer target pseudovirus preparation.

Fig1. Schematic of VSV Pseudovirus Packaging Principle

2. Service Offerings

Drawing on years of experience with viral packaging platforms, GeneRulor has developed a suite of advanced technologies and reagents for the VSV pseudovirus system, substantially improving recombinant VSV packaging titer, purity, reproducibility, and stability. We offer a one-stop VSV pseudovirus solution encompassing:

• Custom pseudovirus construction: client-specified envelope proteins packaged on request

• Pseudovirus packaging & production: high-titer preparations with full QC verification

• Neutralization assay services: reporter gene-based readouts (luciferase or GFP)

• Technical support & data analysis

Fig2. VSV Pseudovirus One-Stop Service Workflow

3. Available Pseudovirus Types

• VSV pseudovirus bearing native VSV-G envelope protein

• VSV pseudovirus pseudotyped with heterologous envelope proteins (including highly pathogenic viruses)

• VSV backbone-based chimeric pseudovirus (including highly pathogenic viruses)

• Bald VSV pseudovirus lacking envelope protein (for use as negative control)

4. Application Areas

• Vaccine development: assessment of vaccine-elicited neutralizing antibody titers

• Antiviral drug screening: evaluation of antiviral compounds and neutralizing antibody efficacy

• Viral mechanism research: investigation of viral entry, membrane fusion, and receptor-binding mechanisms

• Diagnostic development: optimization of serological detection methods (e.g., as surrogate assays for ELISA-based approaches)

5. Technical Advantages

• High biosafety: Pseudoviruses are replication-incompetent, substantially reducing biosafety risk and enabling work under BSL-2 containment.

• High transduction efficiency: The VSV vector platform provides broad host-range tropism and consistently high titers.

• Flexible customization: Supports packaging of diverse viral envelope proteins, including HIV, influenza, Ebola, SARS-CoV-2, and more.

• Stable and reproducible: Optimized for high-throughput screening applications, with excellent inter-assay reproducibility and high data reliability.

References

[1] Whitt, M. A. Generation of VSV pseudotypes using recombinant ΔG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses. Journal of Virology, 2010, 84(2): 844–856.

[2] Langland J, Jacobs B, Wagner CE et al. Antiviral activity of metal chelates of caffeic acid and similar compounds towards herpes simplex, VSV-Ebola pseudotyped and vaccinia viruses. Antiviral Research, 2018 Dec; 160: 143–150.