Rapid determination of HIV-1 mutant frequencies and mutation spectra using an mCHERRY/EGFP dual-reporter viral vector

Jonathan M.O. Rawson, Christine L. Clouser, Louis M. Mansky

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

The high mutation rate of human immunodeficiency virus type-1 (HIV-1) has been a pivotal factor in its evolutionary success as a human pathogen, driving the emergence of drug resistance, immune system escape, and invasion of distinct anatomical compartments. Extensive research has focused on understand­ing how various cellular and viral factors alter the rates and types of mutations produced during viral rep­lication. Here, we describe a single-cycle dual-reporter vector assay that relies upon the detection of mutations that eliminate either expression of mCherry or enhanced green fluorescent protein (EGFP). The reporter-based method can be used to efficiently quantify changes in mutant frequencies and mutation spectra that arise due to a variety of factors, including viral mutagens, drug resistance mutations, cellular physiology, and APOBEC3 proteins.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages71-88
Number of pages18
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1354
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
This work was supported by NIH grant R01 GM105876. J.M.O.R. was supported by NIH grant T32 AI083196 and a University of Minnesota Doctoral Dissertation Fellowship. We are grateful to Cavan Reilly and James Hodges for advice on statistical procedures.

Publisher Copyright:
© Springer Science+Business Media New York 2016.

Keywords

  • Diversity
  • Evolution
  • Lentivirus
  • Mutagenesis
  • Retroviral vector
  • Retrovirus
  • Reverse transcription

Fingerprint

Dive into the research topics of 'Rapid determination of HIV-1 mutant frequencies and mutation spectra using an mCHERRY/EGFP dual-reporter viral vector'. Together they form a unique fingerprint.

Cite this