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A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion

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

DOI:
10.3791/58074-v

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07:22 min

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August 14, 2018

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, , , , ,
1Division of Infectious Diseases, Department of Medicine,Icahn School of Medicine at Mount Sinai, Immunology Institute, 2Department of Biology,New Jersey City University

Capítulos

  • 00:04Título
  • 00:54Preparation of Target Cells for Cell-to-Cell Virus Transmission
  • 02:11Preparation of Donor Cells
  • 03:56Co-culture
  • 05:06Fluorescence-activated Cell Sorting
  • 05:35Results: Identification of a Novel HIV-1 Viral Membrane Fusion Inhibitor
  • 06:49Conclusion

Summary

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We describe a cell-based assay to report on HIV-1 fusion via the expression of green fluorescent protein detectable by flow cytometry or fluorescence microscopy. It can be used to test inhibitors of viral entry (specifically at the fusion step) in cell-free and cell-to-cell infection systems.

Tags

Keywords: Cre-LoxViral Membrane FusionHIV-1Drug DiscoveryHigh-throughput ScreeningHIV Fusion InhibitorsBiosafetyJurkat RG Reporter CellsPuromycinCell-to-cell Virus TransmissionGag-iCre DNAElectroporationFicoll

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