A new generation of functional tagged proteins for hiv fluorescence imaging

João I. Mamede, Joseph Griffin, Stéphanie Gambut, Thomas J. Hope

Research output: Contribution to journalArticlepeer-review

Abstract

During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes to follow individual virus particles while they progress throughout infection. The inclusion of such fusion motifs or surrogates frequently disrupts viral infectivity or results in a change of the wild-type phenotype. Here, we detail the construction and functional characterization of two new constructs where we fused fluorescent proteins to the N-terminus of HIV-1 Integrase. In the first, IN is recruited into assembling particles via a codon optimized Gag to complement other viral constructs, while the second is fused to a Gag-Pol expression vector fully capable of integration. Our data shows that N-terminal tagged IN is functional for integration by both recovery of integration of catalytically inactive IN and by the successful infectivity of viruses carrying only labeled IN. These tools will be important to study the individual behavior of viral particles and associate such behavior to infectivity.

Original languageEnglish (US)
Article number386
JournalViruses
Volume13
Issue number3
DOIs
StatePublished - Mar 2021

Keywords

  • Fluorescent HIV
  • HIV early-steps
  • Integration competent tagged viruses

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

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