Taz protects aged hematopoietic stem cells from exhaustion [scRNA-seq]

We report here the analysis of young and aged hematopoietic stem and progenitor cells (HSPCs) by transcriptional profiling (bulk RNA-Seq, CITE-Seq) and chromatin profiling (ATAC-Seq, scATAC-Seq). We use here a Clca3a1 as a surface marker to further fractionate HSPCs. Young hematopoietic stem cells (HSCs) were isolated by flow cytometry, transduced ex vivo with different shRNA constructs (shRen, shPU1 No1, shPU1 No2) and transplanted into lethally irradiated mice. Two months after transplantation, transduced (GFP+)HSPCs were isolated by flow cytometry. The different samples were pooled and the shRNA per cell was identified by a specific dialout PCR (amplifying the mirE shRNA construct together with the 10x cell barcode) to link the cell barcodes with the the different shRNA constructs. Here mirE_R1 contains the UMI and the cell barcode whereas mirE_R2 contains the shRNA sequence. Examination of transcriptional and chromatin changes during aging of HSPCs.