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

doi:10.1007/978-1-4939-3046-3_6

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

Basic Sciences

Research output: Chapter in Book/Report/Conference proceeding › Chapter

6 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 language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	71-88
Number of pages	18
DOIs	https://doi.org/10.1007/978-1-4939-3046-3_6
State	Published - 2016

Publication series

Name	Methods in Molecular Biology
Volume	1354
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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/978-1-4939-3046-3_6

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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.",

keywords = "Diversity, Evolution, Lentivirus, Mutagenesis, Retroviral vector, Retrovirus, Reverse transcription",

author = "Rawson, {Jonathan M.O.} and Clouser, {Christine L} and Mansky, {Louis M.}",

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: {\textcopyright} Springer Science+Business Media New York 2016.",

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doi = "10.1007/978-1-4939-3046-3_6",

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AU - Rawson, Jonathan M.O.

AU - Clouser, Christine L

AU - Mansky, Louis M.

N1 - 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.

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - Diversity

KW - Evolution

KW - Lentivirus

KW - Mutagenesis

KW - Retroviral vector

KW - Retrovirus

KW - Reverse transcription

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M3 - Chapter

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AN - SCOPUS:84952801903

T3 - Methods in Molecular Biology

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BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

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

Abstract

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Bibliographical note

Keywords

UN SDGs

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