RNA-seq transcriptonal profiling in human primary adult erythroid progenitor cells (ProEs) after shRNA-mediated depletion of TFAM and PHB2 expression

The developing erythroid cells require highly coordinated gene expression and metabolism. By comparing the proteomic and transcriptomic changes in human hematopoietic stem/progenitor cells (HSPCs) and lineage-committed erythroid progenitors (ProEs), and uncover pathways related to mitochondrial biogenesis enhanced through post-transcriptional regulation. Two principal mitochondrial factors TFAM and PHB2 are tightly regulated at the protein level and indispensable for mitochondria and erythropoiesis. To determine the role of TFAM and PHB2 in mitochondrial function during erythroid development, we employed shRNA-mediated depeltion of TFAM and PHB2 expression in differentiating erythroid cells, and performed RNA-seq transcriptional profiling analysis. Overall design: Human primary adult erythroblasts were generated ex vivo from CD34+ hematopoietic stem/progenitor cells (HSPCs) using a serum-free two-phase liquid culture system. CD34+ HSPCs were transduced with lentiviruses containing shRNAs against TFAM, PHB2, or the non-targeting (shNT) control, selected and differentiated to proerythroblasts (ProEs). Cells were harvested at day 5 of erythroid differentiation and total RNA were extracted for RNA-seq analysis.