Overexpression Of miR-451 Induces the Differentiation of Murine Embryonic Stem Cells (MESCs) into Erythroid Lineage

BACKGROUND MiRNAs can play significant roles in growth by targeting the messages of protein-coding genes for the cleavage or suppression of productive translation. MiR-451 is existing in mature circulating red blood cells. Recent studies have shown that miR-451 have key roles in erythropoiesis. METHODS In this experimental study, MESCs were infected with lentiviruses containing pCDH-miR-451. Erythroid differentiation was estimated using quantitative RT-PCR for transcriptional factors (GATA-1, KLF-1, EPOR) and hemoglobin chains (α, β, γ , ε and ζ) genes and flow cytometery for erythroid surface antigens (TER-119 and CD235a). CFU assay was also performed on days 14th and 21st. RESULTS Mature miR-451 expression level increased by 3.434 fold compared to the untreated MESCs on day 4 post transfection (p< 0.001). MiR-451up-regulation could induce transcriptional factors (GATA-1, KLF-1, EPOR) and hemoglobin chains (α, β, γ, ε and ζ) genes expression in mESCs (p< 0.001) and have strong correlation with the appearance of CD235a and Ter-119 markers on these cells (by 13.084 and 13.327 fold, respectively) (p< 0.05). Moreover, MESCs that were treated with pCDH-miR-451 led to a significant raise of CFU-E colonies (by 5.2-fold) compared to untreated control group (p< 0.05). CONCLUSTION Our results showed that overexpression Of miR-451 is essential for erythroid chromatin condensation. Expression of miR-451 has the potential role in the production of artificial RBCs without the presence of any stimulatory cytokines.

MicroRNAs, miR-451, Murine Embryonic Stem Cells (MESCs), Erythropoiesis