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

3 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 - Jan 1 2016

Publication series

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

Keywords

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

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    Rawson, J. M. O., Clouser, C. L., & Mansky, L. M. (2016). Rapid determination of HIV-1 mutant frequencies and mutation spectra using an mCHERRY/EGFP dual-reporter viral vector. In Methods in Molecular Biology (pp. 71-88). (Methods in Molecular Biology; Vol. 1354). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3046-3_6