LAUSR

Significance Gene regulatory logic reflects the occupancy of cis elements by transcription factors and the configuration of promoters and enhancers. As the majority of genome-wide analyses have focused on adult cells, scant attention has been paid to embryonic cells, other than embryonic stem cells. Focusing on genome-wide comparative analyses of two stages of erythroblasts, we discovered that regulation of embryonic-specific genes is promoter-centric through Gata1, whereas adult-specific control is combinatorial enhancer-driven and requires Myb, which is confirmed by increased enhancer–promoter interactions of adult specific genes. Extending genome-wide comparative analyses more broadly to available datasets of diverse mouse and human cells and tissues, we conclude that the progressively increased enhancer dependence of cell-type–specific genes with developmental age is conserved during development. How overall principles of cell-type–specific gene regulation (the “logic”) may change during ontogeny is largely unexplored. We compared transcriptomic, epigenomic, and three-dimensional (3D) genomic profiles in embryonic (EryP) and adult (EryD) erythroblasts. Despite reduced chromatin accessibility compared to EryP, distal chromatin of EryD is enriched in H3K27ac, Gata1, and Myb occupancy. EryP-/EryD-shared enhancers are highly correlated with red blood cell identity genes, whereas cell-type–specific regulation employs different cis elements in EryP and EryD cells. In contrast to EryP-specific genes, which exhibit promoter-centric regulation through Gata1, EryD-specific genes rely more on distal enhancers for regulation involving Myb-mediated enhancer activation. Gata1 HiChIP demonstrated an overall increased enhancer–promoter interactions at EryD-specific genes, whereas genome editing in selected loci confirmed distal enhancers are required for gene expression in EryD but not in EryP. Applying a metric for enhancer dependence of transcription, we observed a progressive reliance on cell-specific enhancers with increasing ontogenetic age among diverse tissues of mouse and human origin. Our findings highlight fundamental and conserved differences at distinct developmental stages, characterized by simpler promoter-centric regulation of cell-type–specific genes in embryonic cells and increased combinatorial enhancer-driven control in adult cells.