Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Systematic discovery of pro- and anti-HIV host factors in primary human CD4+ T cells.

Cell·2026
Same author

CGM23 corresponds to a pan-coronavirus lipopeptide inhibitor potently inhibiting virion fusion.

iScience·2026
Same author

Multiomic analysis of ART-interruption cohorts identifies cell-extrinsic and -intrinsic mechanisms driving lymphocyte-mediated control of HIV rebound.

Immunity·2026
Same author

Pregnancy and lactation induce distinct immune responses to COVID-19 booster vaccination and SARS-CoV-2 breakthrough infection.

JCI insight·2025
Same author

Anatomical, subset, and HIV-dependent expression of viral sensors and restriction factors.

Cell reports·2025
Same author

Fibrin drives thromboinflammation and neuropathology in COVID-19.

Nature·2024

Related Experiment Video

Updated: Mar 16, 2026

A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion
07:22

A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion

Published on: August 14, 2018

7.0K

HIV-1 Fusion Assay.

Marielle Cavrois1, Jason Neidleman1, Warner C Greene1

  • 1GIVI, Gladstone Institute, San Francisco, USA.

Bio-Protocol
|August 16, 2016
PubMed
Summary

This HIV-1 fusion assay tracks viral entry by monitoring the transfer of a BlaM-Vpr protein chimera into target cells. The assay uses a fluorescent substrate that changes color upon cleavage by the chimera, indicating successful viral fusion.

Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Human Immunodeficiency Virus type 1 (HIV-1) infection begins with viral entry into host cells.
  • Understanding the mechanisms of HIV-1 fusion is crucial for developing effective antiviral therapies.
  • Existing assays may not capture all stages of the fusion process.

Purpose of the Study:

  • To present a novel HIV-1 fusion assay.
  • To measure all steps of the HIV-1 life cycle up to and including viral fusion.
  • To provide a quantifiable method for assessing viral entry.

Main Methods:

  • Incorporation of a beta-lactamase Vpr (BlaM-Vpr) protein chimera into HIV-1 virions.
  • Co-incubation of BlaM-Vpr-containing virions with target cells loaded with the fluorescent substrate CCF2.

More Related Videos

Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
07:10

Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay

Published on: September 14, 2014

14.8K
Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection
07:24

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Published on: October 29, 2020

3.3K

Related Experiment Videos

Last Updated: Mar 16, 2026

A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion
07:22

A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion

Published on: August 14, 2018

7.0K
Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
07:10

Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay

Published on: September 14, 2014

14.8K
Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection
07:24

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Published on: October 29, 2020

3.3K
  • Detection of BlaM-Vpr chimera transfer via enzymatic cleavage of CCF2, resulting in a blue fluorescence emission.
  • Quantification of fusion events using flow cytometry.

    • Main Results:

    • The assay successfully measures virion fusion by detecting the transfer of BlaM-Vpr into target cells.
    • Cleavage of CCF2 by BlaM-Vpr results in a distinct shift in fluorescence from green to blue.
    • Flow cytometry allows for sensitive and quantitative detection of fusion events.

    Conclusions:

    • The developed HIV-1 fusion assay is a reliable method for studying viral entry.
    • This assay provides a comprehensive measure of the fusion process in the HIV-1 life cycle.
    • The assay's sensitivity and quantitative nature make it valuable for antiviral drug screening and virology research.