Open AccessReptin Regulates Pluripotency of Embryonic Stem Cells and Somatic Cell Reprogramming Through Oct4‐Dependent Mechanism
Author(s) -
Do Eun kyoung,
Cheon Hyo Cheon,
Jang Il Ho,
Choi Eun Jung,
Heo Soon Chul,
Kang Kyung Taek,
Bae Kwang Hee,
Cho Yee Sook,
Seo Jeong Kon,
Yoon Jong Hyuk,
Lee Taehoon G.,
Kim Jae Ho
Publication year - 2014
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.1827
Subject(s) - reprogramming , induced pluripotent stem cell , biology , homeobox protein nanog , embryonic stem cell , sox2 , microbiology and biotechnology , rex1 , somatic cell , stem cell , cell potency , small hairpin rna , gene knockdown , genetics , cell , cell culture , gene
Oct4 has been implicated in regulation of pluripotency in embryonic stem cells (ESCs) and reprogramming of somatic cells into induced pluripotent stem cells. However, the molecular mechanisms involved in Oct4‐dependent regulation of pluripotency and reprogramming have not been clear. To gain insight into the mechanism of regulation of Oct4‐mediated self‐renewal of ESCs and reprogramming of somatic cells, we attempted to identify Oct4‐binding proteins using affinity purification and mass spectrometry. We identified Reptin, a key component of ATP‐dependent chromatin remodeling complexes, as an Oct4‐binding protein. Depletion of endogenous Reptin using lentiviral short hairpin RNA (shRNA) led to a decrease in the number and size of alkaline phosphatase‐positive colonies of mouse ESCs. In addition, shRNA‐mediated silencing of Reptin resulted in decreased expression of pluripotency‐specific marker genes, including Oct4, Sox2, Nanog, and SSEA‐1. Results of the Oct4 reporter assay showed synergism between Oct4 and Reptin, and depletion of endogenous Reptin abolished Oct4 transcriptional activity. Results of a chromatin immunoprecipitation assay showed the overlapping interaction of Reptin and Oct4 to CR4 in the Oct4 enhancer in ESCs. Knockdown of Reptin using shRNA suppressed the reprogramming of mouse embryonic fibroblasts to induced pluripotent stem cells, whereas overexpression of Reptin resulted in enhanced efficiency of induced pluripotent stem cell generation. These results strongly suggest that Reptin plays a key role in maintaining the pluripotency of ESCs and in establishing the pluripotency during reprogramming of somatic cells by regulation of Oct4‐mediated gene regulation. S tem C ells 2014;32:3126–3136