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A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion
Published on: August 14, 2018
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Transient Transfection-based Fusion Assay for Viral Proteins
Melina Vallbracht1, Christina Schröter1, Barbara G Klupp1
1Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Bio-Protocol
|August 30, 2021
Summary
This study details a new fusion assay for studying enveloped virus entry. The optimized protocol quantifies cell-cell fusion mediated by pseudorabies virus (PrV) glycoproteins.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Membrane fusion is essential for enveloped virus entry and cell-to-cell spread.
- Alphaherpesviruses like pseudorabies virus (PrV) utilize specific glycoproteins for membrane fusion.
Purpose of the Study:
- To develop and optimize a transient transfection-based fusion assay.
- To quantify cell-cell fusion induced by the minimal PrV glycoprotein set (gB, gH, gL).
Main Methods:
- Transient transfection of cells with PrV glycoproteins gB, gH, and gL.
- Development of an infection-free system to study fusion.
- Quantification of cell-cell fusion events.
Main Results:
- An optimized protocol for a fusion assay was established.
- The assay successfully quantifies fusion mediated by PrV glycoproteins.
- This system allows detailed study of the viral fusion mechanism.
Conclusions:
- The developed assay provides a robust method to study alphaherpesvirus-mediated membrane fusion.
- This infection-free system is valuable for dissecting viral entry mechanisms.
- Understanding PrV glycoprotein function advances knowledge of enveloped virus fusion.
