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 language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Publisher||Humana Press Inc.|
|Number of pages||18|
|State||Published - 2016|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding 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.
© Springer Science+Business Media New York 2016.
- Retroviral vector
- Reverse transcription