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

Methods : a Companion to Methods in Enzymology
Benjamin DaleBenjamin K Chen

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.

References

Aug 6, 2003·The Journal of Cell Biology·Owen PornillosWesley I Sundquist

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Citations

Jan 21, 2014·Nature Chemical Biology·Thomas A Rando
Mar 19, 2014·Developmental and Comparative Immunology·Hayley DirscherlJill L O de Jong
May 7, 2014·AIDS Research and Human Retroviruses·Ashwanth Christopher FrancisAnna Cereseto
Nov 22, 2014·The Journal of Infectious Diseases·Deborah J Anderson
Jan 11, 2013·Journal of the Royal Society, Interface·Tais A P F DollPeter Burkhard
Jan 22, 2011·Methods : a Companion to Methods in Enzymology·Melanie Ott
Jan 29, 2013·Journal of Hepatology·Luke W MeredithJane A McKeating
May 28, 2013·Journal of Virological Methods·Sonia Gutiérrez-GranadosMaría Mercedes Segura
May 4, 2016·The Journal of Physical Chemistry. B·Bhaven MistryTom Chou
Nov 17, 2017·Applied Microbiology and Biotechnology·João VidigalAna P Teixeira
Sep 13, 2019·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Ryan P McNamara, Dirk P Dittmer
Feb 5, 2021·Journal of Molecular Biology·Yisong DengJames R Williamson

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