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High-throughput single-cell sequencing for retroviral reservoir characterization

Lauren E Droske, Stephen D. Shank, View ORCID ProfileMelanie N Cash, Sergei L Kosakovsky Pond, Marco Salemi, Brittany Rife Magalis
doi: https://doi.org/10.1101/2022.05.04.490630
Lauren E Droske
aUniversity of Florida, Department of Pathology, Immunology, and Laboratory Medicine, Gainesville, 32610, USA
bUniversity of Florida, Emerging Pathogens Institute, Gainesville, 32610, USA
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Stephen D. Shank
cTemple University, Department of Biology, Philadelphia, 19122, USA
dTemple University, Institute for Genomics and Evolutionary Medicine, Philadelphia, 19122, USA
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Melanie N Cash
aUniversity of Florida, Department of Pathology, Immunology, and Laboratory Medicine, Gainesville, 32610, USA
bUniversity of Florida, Emerging Pathogens Institute, Gainesville, 32610, USA
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  • ORCID record for Melanie N Cash
Sergei L Kosakovsky Pond
cTemple University, Department of Biology, Philadelphia, 19122, USA
dTemple University, Institute for Genomics and Evolutionary Medicine, Philadelphia, 19122, USA
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Marco Salemi
aUniversity of Florida, Department of Pathology, Immunology, and Laboratory Medicine, Gainesville, 32610, USA
bUniversity of Florida, Emerging Pathogens Institute, Gainesville, 32610, USA
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  • For correspondence: brittany.rife@ufl.edu salemi@pathology.ufl.edu
Brittany Rife Magalis
aUniversity of Florida, Department of Pathology, Immunology, and Laboratory Medicine, Gainesville, 32610, USA
bUniversity of Florida, Emerging Pathogens Institute, Gainesville, 32610, USA
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  • For correspondence: brittany.rife@ufl.edu salemi@pathology.ufl.edu
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ABSTRACT

During the course of infection, human immunodeficiency virus (HIV) maintains a stably integrated reservoir of replication-competent proviruses within the host genome that are unaffected by antiretroviral therapy. Curative advancements rely heavily on targeting the reservoir, though determinants of its evolutionary origins remain ill-supported through current strategies and are often limited by sample variety. Here, we describe a single-cell deoxyribonucleic acid sequencing (scDNA-seq) method, optimized for sequencing of proviral and host DNA from a treatment-interrupted HIV animal model. We report its benefits for improving viral reservoir resolution to support critical evolutionary events otherwise considered unreliable using traditional viral envelope gene signal alone, as well as comparative advantages to existing near-full-length genome sequencing methods. Given the variety of proviral characteristics that may influence viral rebound, scDNA-seq holds immense value in its ability to streamline many of the present-day applications available in viral reservoir studies, such as integration status and putative replication competency.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 04, 2022.
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High-throughput single-cell sequencing for retroviral reservoir characterization
Lauren E Droske, Stephen D. Shank, Melanie N Cash, Sergei L Kosakovsky Pond, Marco Salemi, Brittany Rife Magalis
bioRxiv 2022.05.04.490630; doi: https://doi.org/10.1101/2022.05.04.490630
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High-throughput single-cell sequencing for retroviral reservoir characterization
Lauren E Droske, Stephen D. Shank, Melanie N Cash, Sergei L Kosakovsky Pond, Marco Salemi, Brittany Rife Magalis
bioRxiv 2022.05.04.490630; doi: https://doi.org/10.1101/2022.05.04.490630

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