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Distinct Enhancer Activity of Oct4 in Naive and Primed Mouse Pluripotency.
- Source :
-
Stem cell reports [Stem Cell Reports] 2016 Nov 08; Vol. 7 (5), pp. 911-926. Date of Electronic Publication: 2016 Oct 27. - Publication Year :
- 2016
-
Abstract
- Naive and primed pluripotent stem cells (PSCs) and germ cells express the Oct4 gene. The Oct4 gene contains two cis-regulatory elements, the distal enhancer (DE) and proximal enhancer (PE), which differentially control Oct4 expression in a cell-type-specific and stage-specific manner. Here, we generated double transgenic mice carrying both Oct4-ΔPE-GFP and Oct4-ΔDE-tdTomato (RFP), enabling us to simultaneously monitor the activity of DE and PE. Oct4 expression is stage-specifically regulated by DE and PE during embryonic and germ cell development. Using this dual reporter system, we successfully cultured pure populations of naive (GFP <superscript>+</superscript> RFP <superscript>-</superscript> ) and primed (GFP <superscript>-</superscript> RFP <superscript>+</superscript> ) PSCs. We found that GFP <superscript>+</superscript> RFP <superscript>-</superscript> cells were metastable (not naive) in serum-containing medium; stable naive pluripotent cells were observed in medium containing two inhibitors (Meki and GSKi) but lacked serum. Finally, we suggest that the activity of Oct4 DE and PE is regulated by the repressive histone marks and DNA methylation in a cell-type-specific manner.<br /> (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Animals
Blastocyst cytology
Blastocyst metabolism
Cell Differentiation genetics
Cluster Analysis
DNA Methylation
Embryonic Development
Embryonic Stem Cells cytology
Embryonic Stem Cells metabolism
Gene Expression Profiling
Gene Expression Regulation, Developmental
Genes, Reporter
Histones metabolism
Mice
Mice, Transgenic
Regulatory Sequences, Nucleic Acid
Transcriptome
Enhancer Elements, Genetic
Octamer Transcription Factor-3 genetics
Octamer Transcription Factor-3 metabolism
Pluripotent Stem Cells cytology
Pluripotent Stem Cells metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2213-6711
- Volume :
- 7
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Stem cell reports
- Publication Type :
- Academic Journal
- Accession number :
- 28157483
- Full Text :
- https://doi.org/10.1016/j.stemcr.2016.09.012