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. 2013 Nov;11(3):1299-313.
doi: 10.1016/j.scr.2013.09.001. Epub 2013 Sep 13.

MicroRNA-137 Represses Klf4 and Tbx3 During Differentiation of Mouse Embryonic Stem Cells

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MicroRNA-137 Represses Klf4 and Tbx3 During Differentiation of Mouse Embryonic Stem Cells

Ke Jiang et al. Stem Cell Res. .

Abstract

MicroRNA-137 (miR-137) has been shown to play an important role in the differentiation of neural stem cells. Embryonic stem (ES) cells have the potential to differentiate into different cell types including neurons; however, the contribution of miR-137 in the maintenance and differentiation of ES cells remains unknown. Here, we show that miR-137 is mainly expressed in ES cells at the mitotic phase of the cell cycle and highly upregulated during differentiation. We identify that ES cell transcription factors, Klf4 and Tbx3, are downstream targets of miR-137, and we show that endogenous miR-137 represses the 3' untranslated regions of Klf4 and Tbx3. Transfection of ES cells with mature miR-137 RNA duplexes led to a significant reduction in cell proliferation and the expression of Klf4, Tbx3, and other self-renewal genes. Furthermore, we demonstrate that increased miR-137 expression accelerates differentiation of ES cells in vitro. Loss of miR-137 during ES cell differentiation significantly impeded neuronal gene expression and morphogenesis. Taken together, our results suggest that miR-137 regulates ES cell proliferation and differentiation by repressing the expression of downstream targets, including Klf4 and Tbx3.

Keywords: 3′ untranslated region; 3′UTR; 4,6-diamidino-2-phenylindole dihydrochloride; DAPI; EB; ES cells; LIF; LSD1; NC; PBS; RT; SSC; Tuj1; ddPCR; droplet digital PCR; embryoid body; embryonic stem cells; leukemia inhibitory factor; lysine specific demethylase 1; miRNA; microRNA; neuronal class III β-tubulin 3; neuronally differentiated ES cells; phosphate buffered saline; qPCR; quantitative real-time polymerase chain reaction; reverse transcription; saline-sodium citrate.

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