Using Barcoded HIV Ensembles (B-HIVE) for Single Provirus Transcriptomics.

Area of Science:

• Virology
• Immunology
• Molecular Biology

Background:

• The persistent latent HIV reservoir is a major obstacle to curing AIDS.
• Latent HIV-infected cells evade immune surveillance and antiretroviral therapies.
• Effective treatment strategies necessitate methods to track latent proviruses.

Purpose of the Study:

• To introduce a novel genome-wide technology, Barcoded HIV Ensembles (B-HIVE), for measuring HIV expression at the single provirus level.
• To enable the quantification and characterization of individual viral transcripts originating from single proviruses.

Main Methods:

• B-HIVE utilizes DNA barcodes to tag the HIV genome, allowing for the tracing of viral transcripts produced by single proviruses.
• This high-throughput method quantifies thousands of individual viral transcripts per infection cycle.
• The technique can be applied across various experimental conditions to assess single provirus responses to treatments.

Main Results:

• B-HIVE successfully identifies and quantifies HIV expression at the single provirus level.
• The method distinguishes between active, latent, and low-level expressed proviruses within a cell population.
• It provides unprecedented insights into the heterogeneity of single provirus expression.

Conclusions:

• B-HIVE is a powerful tool for understanding HIV latency and expression dynamics.
• This technology facilitates the evaluation of novel therapeutic interventions targeting the latent HIV reservoir.
• B-HIVE offers a new avenue for advancing research towards an AIDS cure.