Viral Pseudotype Production
Viral Pseudotype Production - Related Videos
11:08
Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles
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Cited by 15 •
2012
Lentiviral expression vectors are the most effective vehicles for stably expressing different effector molecules or reporter constructs in dividing and non-dividing mammalian cells and whole organisms. Here we provide a protocol on how to package lentivector expression constructs in pseudoviral particles and to transduce target cells using the pseudoviral particles.
08:35
Production and Titering of Recombinant Adeno-associated Viral Vectors
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Cited by 149 •
2011
Recombinant adeno-associated virus (rAAVs) vectors are becoming increasingly valuable for in vivo studies in animals. We describe how rAAVs can be produced in the laboratory and how these vectors can be titered to give an accurate reading of the number of infectious particles produced.
08:32
Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production
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Cited by 2 •
2014
We describe a protocol to measure the antiviral cytokine production in mice infected with a model herpesvirus, murine gamma herpesvirus 68 (γHV68) that is closely-related to human Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV). Utilizing genetically modified mouse strains and mouse embryonic fibroblasts (MEFs), we assessed the antiviral cytokine production both in vivo and ex vivo. “Reconstituting” the expression of innate immune components in knockout embryonic...
02:57
Assessing Antiviral Cytokine Production in Response to Viral Infection
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2026
Source: Zhang, J., et al. Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production. J. Vis. Exp. (2014)This video demonstrates the process of infecting murine fibroblast cells with herpesvirus and monitoring the resulting antiviral cytokine release and gene expression. It outlines key steps, including viral infection, media collection for cytokine quantification, and cell harvesting for gene expression analysis.
05:49
Pseudotyped Viruses As a Molecular Tool to Monitor Humoral Immune Responses Against SARS-CoV-2 Via Neutralization Assay
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2023
Pseudotyped viruses (PVs) are replication-defective virions that are used to study host-virus interactions under safer conditions than handling authentic viruses. Presented here is a detailed protocol that shows how SARS-CoV-2 PVs can be used to test the neutralizing ability of patients' serum after COVID-19 vaccination.
FAQs
The engineered particle combines two functionally distinct components: the structural core supplies the particle framework, while the selected envelope protein supplies surface features that interact with host cells. This division lets investigators examine entry properties associated with the displayed glycoprotein without treating the particle as a complete representation of the original virus.
Keeping entry function separate from the genome responsible for replication is central to the system. The resulting particles can enter selected host cells while generally not undergoing productive infection. This separation supports controlled studies of viral entry and reduces reliance on replication-competent pathogens in immunology and infection research.
Researchers can compare particles displaying different viral envelope proteins and observe whether they enter particular host cells. Differences in entry patterns provide information about tropism, meaning the range of cells a viral surface can target. The approach therefore links envelope variation with host-cell selection while keeping the particle framework more consistent.
Host-cell receptor usage can be investigated by examining which cells are entered by particles bearing a particular envelope protein. Because the envelope mediates the relevant surface interaction, changes in entry across cell types can help relate viral glycoproteins to receptor-dependent targeting. This makes pseudotypes useful for dissecting entry pathways in a controlled setting.
The design requires a structural viral core paired with an envelope protein selected from the virus or variant under study. The core and displayed glycoprotein must remain functionally distinct so that the particle presents the desired entry properties without relying on the original virus’s complete replication program. This arrangement supports focused downstream measurements.
Particles displaying different envelope variants can be evaluated side by side for their ability to enter selected host cells or to be blocked by antibodies or serum. Comparing these outcomes helps researchers identify changes in tropism, entry behavior, or immune recognition. The same platform can therefore connect envelope variation with both infection-related and immunological readouts.
Pseudotypes provide a platform for evaluating antibody or serum neutralization by testing whether immune factors interfere with envelope-mediated entry. Researchers can also use the system to assess candidate antiviral strategies that target this step. In immunology and infection studies, these measurements help compare immune responses and identify approaches that limit viral entry.