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An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
Published on: June 1, 2018
DNA hypermethylation enhanced telomerase reverse transcriptase expression in human-induced pluripotent stem cells
Ken Takasawa1, Yoshikazu Arai1, Mayu Yamazaki-Inoue2
1Laboratory of Veterinary Biochemistry and Molecular Biology, Graduate School of Medicine and Veterinary Medicine/Faculty of Agriculture, University of Miyazaki, 1-1 Gakuen-Kibanadai-Nishi, Miyazaki, 889-2192, Japan.
DNA methylation regulates TERT gene expression during induced pluripotent stem cell (iPSC) reprogramming. A specific TERT promoter region becomes methylated in iPSCs, enhancing TERT transcription and gene expression.
Area of Science:
- Epigenetics
- Stem Cell Biology
- Molecular Biology
Background:
- Induced pluripotent stem cells (iPSCs) are generated by reprogramming somatic cells.
- Stem cell marker genes like OCT-4, NANOG, SALL4, and TERT are induced during reprogramming.
- DNA methylation regulates OCT-4, NANOG, and SALL4 expression via promoter hypomethylation.
Purpose of the Study:
- To investigate the epigenetic regulation of the human TERT gene during iPSC reprogramming.
- To identify regulatory elements and mechanisms controlling TERT gene expression.
Main Methods:
- Identification of a differentially methylated region (DMR) in the TERT promoter.
- Region-specific methylated-promoter assays.
- Analysis of Lamin B1 binding to the TERT-DMR.
Main Results:
- A novel TERT-DMR was identified in a distal promoter region.
- The TERT-DMR was highly methylated in iPSCs and hypomethylated in parental somatic cells.
- Methylated TERT-DMR up-regulated TERT promoter activity in iPSCs.
- Lamin B1 accumulated at the TERT-DMR in iPSCs.
Conclusions:
- DNA methylation at the TERT-DMR enhances TERT transcription during iPSC reprogramming.
- TERT transcription is promoted by DNA methylation via nuclear lamina binding.
- This study reveals a novel role for DNA methylation in TERT gene regulation.
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