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Related Experiment Video

Updated: Mar 2, 2026

High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing
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High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing

Published on: January 22, 2019

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High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay.

Robert W Yucha1, Kristen S Hobbs2, Emily Hanhauser2

  • 1Division of Infectious Diseases, Brigham and Women's Hospital, 75 Francis Street, Boston, MA 02115, United States.

Ebiomedicine
|May 23, 2017
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel assay to measure HIV-1 transcription in single cells. This tool helps understand how latency reversing agents (LRAs) affect HIV reservoirs, aiding eradication research.

Keywords:
Digital PCRHIV reactivationHIV reservoirsHistone deacetylase inhibitorsHuman Immunodeficiency Virus (HIV)Single cell quantification

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Area of Science:

  • Virology
  • Immunology
  • Molecular Biology

Background:

  • Reactivation of latent HIV-1 reservoirs is critical for eradication strategies.
  • The precise impact of latency reversing agents (LRAs) on HIV-1 transcription levels in different cell states remains unclear.

Purpose of the Study:

  • To develop and validate a sensitive assay for quantifying HIV-1 transcriptional activity at the single-cell level.
  • To investigate the effects of specific LRAs (romidepsin and TCR stimulation) on HIV-1 RNA production in CD4+ T cells from infected individuals.

Main Methods:

  • Development of a microfluidic single-cell-in-droplet (scd)PCR assay.
  • Detection of unspliced (us)RNA and multiply spliced (ms)RNA to measure HIV-1 transcriptional activity.
  • Application of the scdPCR assay to CD4+ T cells from HIV-1-infected individuals on antiretroviral therapy.

Main Results:

  • The scdPCR assay can detect and quantify HIV-1 transcriptional activity, including genomic DNA and mRNA, in individual cells.
  • LRAs demonstrated heterogeneous effects, increasing transcription in some cells and reactivating latent cells in others.
  • The assay allows for high-throughput analysis of HIV-1 reactivation in hundreds of thousands of cells.

Conclusions:

  • The developed scdPCR assay provides a powerful tool for studying HIV-1 latency and reactivation dynamics.
  • Findings suggest that LRAs have varied impacts on HIV-1 reservoirs, necessitating personalized approaches for eradication.
  • This assay has the potential to significantly advance research into HIV-1 reservoir eradication strategies.