Tracking and quantitation of fluorescent HIV during cell-to-cell transmission

Benjamin M. Dale, Gregory P. McNerney, Wolfgang Hübner, Thomas R Huser, Benjamin K. Chen

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

The green fluorescent protein (GFP) is a powerful genetic marking tool that has enabled virologists to monitor and track viral proteins during HIV infection. Expression-optimized Gag-GFP constructs have been used to study virus-like particle (VLP) assembly and localization in cell types that are easily transfected. The development of HIV-1 variants carrying GFP within the context of the viral genome has facilitated the study of infection and has been particularly useful in monitoring the transfer of virus between cells following virological synapse formation. HIV Gag-iGFP, a viral clone that contains GFP inserted between the matrix (MA) and capsid (CA) domains of Gag, is the first replication competent molecular clone that generates fluorescent infectious particles. Here, we discuss some methods that exploit HIV Gag-iGFP to quantify cell-to-cell transmission of virus by flow cytometry and to track the proteins during assembly and transmission using live-cell imaging.

Original languageEnglish (US)
Pages (from-to)20-26
Number of pages7
JournalMethods
Volume53
Issue number1
DOIs
StatePublished - Jan 2011

Keywords

  • Flow cytometry
  • Gag
  • GFP
  • HIV
  • Live imaging
  • Virological synapse

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint

Dive into the research topics of 'Tracking and quantitation of fluorescent HIV during cell-to-cell transmission'. Together they form a unique fingerprint.

Cite this