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CG Dinucleotide Removal in Bioluminescent and Fluorescent Reporters Improves HIV-1 Replication and Reporter Gene Expression for Dual Imaging in Humanized Mice.
- Source :
-
Journal of virology [J Virol] 2021 Sep 09; Vol. 95 (19), pp. e0044921. Date of Electronic Publication: 2021 Sep 09. - Publication Year :
- 2021
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Abstract
- Visualizing the transmission and dissemination of human immunodeficiency virus type 1 (HIV-1) in real time in humanized mouse models is a robust tool to investigate viral replication during treatments and in tissue reservoirs. However, the stability and expression of HIV-1 reporter genes are obstacles for long-term serial imaging in vivo . Two replication-competent CCR5-tropic HIV-1 reporter constructs were created that encode either nanoluciferase (nLuc) or a near - infrared fluorescent protein (iRFP) upstream of nef . HIV-1 reporter virus replication and reporter gene expression was measured in cell culture and in humanized mice. While reporter gene expression in vivo correlated initially with plasma viremia, expression decreased after 4 to 5 weeks despite high plasma viremia. The reporter genes were codon optimized to remove cytosine/guanine (CG) dinucleotides, and new CO-nLuc and CO-iRFP viruses were reconstructed. Removal of CG dinucleotides in HIV-1 reporter viruses improved replication in vitro and reporter expression in vivo and ex vivo . Both codon-optimized reporter viruses could be visualized during coinfection and in vivo reporter gene expression during treatment failure preceded detection of plasma viremia. While the dynamic range of CO-iRFP HIV-1 was lower than that of CO-nLuc HIV-1, both viruses could have utility in studying and visualizing HIV-1 infection in humanized mice. IMPORTANCE Animal models are important for studying HIV-1 pathogenesis and treatments. We developed two viruses each encoding a reporter gene that can be expressed in cells after infection. This study shows that HIV-1 infection can be visualized by noninvasive, whole-body imaging in mice with human immune cells over time by reporter expression. We improved reporter expression to reflect HIV-1 replication and showed that two viral variants can be tracked over time in the same animal and can predict failure of antiretroviral therapy to suppress virus.
- Subjects :
- Animals
CD4-Positive T-Lymphocytes virology
Gene Expression
HIV-1 genetics
Humans
Luciferases genetics
Luminescent Measurements
Luminescent Proteins genetics
Mice
Optical Imaging
Viremia
Whole Body Imaging
Dinucleoside Phosphates metabolism
Genes, Reporter
HIV Infections virology
HIV-1 physiology
Virus Replication
Subjects
Details
- Language :
- English
- ISSN :
- 1098-5514
- Volume :
- 95
- Issue :
- 19
- Database :
- MEDLINE
- Journal :
- Journal of virology
- Publication Type :
- Academic Journal
- Accession number :
- 34232063
- Full Text :
- https://doi.org/10.1128/JVI.00449-21