Pseudovirus Production

Pseudovirus production is the laboratory generation of engineered, replication-defective viral particles that mimic selected aspects of a virus without carrying a complete genome capable of sustained replication. It typically involves expressing viral structural and envelope proteins together with a reporter or other cargo, allowing particles to assemble in producer cells and deliver their payload to susceptible target cells through receptor-mediated entry. In biology, these particles provide a controlled platform for studying viral tropism, cell entry, and host-factor requirements while reducing the risks associated with working with fully infectious viruses. They also support neutralization assays, vaccine evaluation, antiviral screening, and analysis of emerging viral variants.

Pseudovirus Production - Related Videos

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JoVE EoE - Viral Growth and Techniques

Production of Pseudovirus Using Plasmid Transfection in Human Cells

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2026

Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)This video demonstrates the production of influenza pseudoviruses in human cells using calcium phosphate-mediated plasmid transfection, enabling safe generation of single-cycle viral particles for downstream applications such as antibody neutralization assays under biosafety level 2 conditions.

Production and Collection of Pseudovirus Particles Using Mammalian Cell Culture

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2026

Source: Jamieson, T. R., et al. Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection. J. Vis. Exp. (2021)This video demonstrates the procedure for infecting mammalian cells with green fluorescent protein (GFP)-encoding pseudovirus, monitoring viral replication and spread by fluorescence microscopy, and collecting virus-containing supernatant for downstream applications.

Assessing Pseudovirus Infection Using Luciferase Reporter Assay

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2026

Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021).This video demonstrates a luciferase-based assay to quantify pseudovirus infection in epithelial cells. The pseudovirus encodes the luciferase gene, which is expressed in infected cells. After washing, and cells are incubated with lysis buffer at an ultra-low temperature, and then thawed to induce lysis.

Quantification of Pseudovirus Particles by Plaque Assay

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2026

Source: Jamieson, T. R., et al., Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection. J. Vis. Exp. (2021)This video describes the virus plaque assay using coronavirus pseudovirus particles. The diluted pseudovirus is added to a monolayer of mammalian cells, followed by incubation with a methylcellulose overlay. Plaques formed by infected cells are counted visually to determine the infectious virus titer.

Determining Optimal Target Cell Density Using a Luciferase-Based Pseudovirus Infection Assay

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2026

Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)This video demonstrates a luciferase-based pseudovirus titration assay to determine the optimal target cell density for maximum infection. By comparing luminescence across cell densities, the method ensures high assay sensitivity and reproducibility for downstream applications.

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