Control samples for study "hiv-seq reveals global host gene expression differences between hiv-transcribing cells from viremic and suppressed people with hiv"

In our study, we aimed to improve the identification of active reservoir cells producing HIV in individuals undergoing antiretroviral therapy (ART). We developed a technique called HIV-Seq to enhance the efficiency of capturing HIV transcripts during single-cell RNA sequencing (scRNAseq) analysis. By applying this technique, we compared the transcriptional features of HIV RNA+ cells before and after ART initiation. Our findings highlight distinct transcriptional signatures of active reservoir cells during viremia and suppression, demonstrating the utility of HIV-Seq in elucidating the mechanisms underlying reservoir persistence during ART. Overall design: CD4? T cells from two HIV-negative donors were subjected to both the standard scRNA-seq protocol and the HIV-seq protocol in parallel. This was done to validate that the addition of HIV-specific primers in the HIV-seq method does not result in false-positive detection of HIV transcripts. Peripheral blood mononuclear cells (PBMCs) were thawed, and CD4? T cells were isolated by negative selection using the EasySep™ Human CD4? T Cell Enrichment Kit (StemCell Technologies). These purified cells were then used to generate gene expression (GEX) libraries following either the standard 10x Genomics 5' scRNA-seq protocol or the modified HIV-seq protocol.