title: Tet and TDG mediate DNA demethylation essential for mesenchymal-to-epithelial transition in somatic cell reprogramming.
abstract: Tet-mediated DNA oxidation is a recently identified mammalian epigenetic modification, and its functional role in cell-fate transitions remains poorly understood. Here, we derive mouse embryonic fibroblasts (MEFs) deleted in all three Tet genes and examine their capacity for reprogramming into induced pluripotent stem cells (iPSCs). We show that Tet-deficient MEFs cannot be reprogrammed because of a block in the mesenchymal-to-epithelial transition (MET) step. Reprogramming of MEFs deficient in TDG is similarly impaired. The block in reprogramming is caused at least in part by defective activation of key miRNAs, which depends on oxidative demethylation promoted by Tet and TDG. Reintroduction of either the affected miRNAs or catalytically active Tet and TDG restores reprogramming in the knockout MEFs. Thus, oxidative demethylation to promote gene activation appears to be functionally required for reprogramming of fibroblasts to pluripotency. These findings provide mechanistic insight into the role of epigenetic barriers in cell-lineage conversion.
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Xiao
authors: X
authors: Hu
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Lei
authors: L
authors: Zhang
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Shi-Qing
authors: SQ
authors: Mao
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Zheng
authors: Z
authors: Li
authors: CAS Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors: Jiekai
authors: J
authors: Chen
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Run-Rui
authors: RR
authors: Zhang
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Hai-Ping
authors: HP
authors: Wu
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Juan
authors: J
authors: Gao
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Fan
authors: F
authors: Guo
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Wei
authors: W
authors: Liu
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Gui-Fang
authors: GF
authors: Xu
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Hai-Qiang
authors: HQ
authors: Dai
authors: Division of Endocrinology, Diabetes, and Hypertension, Department of Medicine and BCMP, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Yujiang Geno
authors: YG
authors: Shi
authors: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China.
authors: Xianlong
authors: X
authors: Li
authors: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China.
authors: Boqiang
authors: B
authors: Hu
authors: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China.
authors: Fuchou
authors: F
authors: Tang
authors: CAS Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors: Duanqing
authors: D
authors: Pei
authors: Shanghai Key Laboratory of Molecular Andrology, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: glxu@sibs.ac.cn.
authors: Guo-Liang
authors: GL
authors: Xu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D045647:DNA Glycosylases / Q000502:physiology*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000502:physiology*; D049308:Dioxygenases; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D044127:Epigenesis, Genetic; D058750:Epithelial-Mesenchymal Transition; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D007124:Immunoenzyme Techniques; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D004268:DNA-Binding Proteins; D035683:MicroRNAs; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; C540839:TET1 protein, mouse; D049308:Dioxygenases; C551914:Tet2 protein, mouse; C551915:Tet3 protein, mouse; D045647:DNA Glycosylases