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RUNX1-Induced DNA Demethylation Drives Hematopoietic Stem and Progenitor Cell Development

ABSTRACT: This study investigates the role of RUNX1’s DNA demethylation function in hematopoietic differentiation, revealing its necessity for generating hematopoietic stem and progenitor cells (HSPCs). Using CRISPR-edited iPSC lines—TADΔ (truncated transactivation domain) and Runt-VP64 (TAD replaced with VP64)—we demonstrated that RUNX1’s ability to induce DNA demethylation is critical for HSPC emergence. While wild-type RUNX1 enabled robust HSPC production (41.1% efficiency), Runt-VP64 and TADΔ severely impaired differentiation (13.5% and 0.024% efficiency, respectively). Methylome analysis linked RUNX1-binding motifs to differentially methylated regions (DMRs) in wild-type cells, absent in mutants, highlighting RUNX1’s role in site-specific demethylation during progenitor cell emergence. These findings establish RUNX1’s DNA demethylation activity as essential for initiating hematopoietic lineage commitment, providing novel insights into epigenetic regulation of hematopoiesis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE300808 | GEO | 2025/07/01

REPOSITORIES: GEO

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RUNX1-Mediated Binding Site Directed DNA Demethylation is a Novel Mechanism of Hematopoietic Gene Activation.

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