Development of a multiplexed approach for isolation and simultaneous protein and transcriptome profiling of HIV-1 protein-containing cells in the brain

National Institute On Drug Abuse

Investigators

Abstract

Funding for this project was obtained late in the fiscal year so scientific progress towards proposed aims has been limited. So far we have acquired experimental supplies and reagents for the project, and recruited three post baccalaureate fellows to work on the project. We also initiated contracts for generation of custom single nucleus RNA sequencing (snRNA-seq) reagents specific to the HIV-1 proteins of interest to support proposed studies. Ongoing work will first validate the new reagents, and then use them to identify HIV-1 infected single cells expressing proteins of interest and uninfected single cells that have taken up these proteins. We will adapt new snRNA-seq pipelines developed in Hope lab to isolate these two rare populations (based on HIV-protein signal) and integrate protein levels with transcriptomic data at single cell resolution. This will provide a comprehensive view of how varying levels of HIV-1 proteins influence CNS cell states beyond for the primary infected and non-infected cell.