Journal
STEM CELLS
Volume 26, Issue 11, Pages 2821-2831Publisher
WILEY
DOI: 10.1634/stemcells.2008-0482
Keywords
Reprogramming; Cell fusion; Pluripotency; Oct4; Xist; Histone deacetylase; Dnmt3a; Tsix
Categories
Funding
- Federal Ministry of Education and Research (BMBF) initiative Cell-Based
- Regenerative Medicine [01GN0539]
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Reactivation of Oct4 gene expression occurs within 2 days of fusion of somatic cells with pluripotent stem cells and within 9 days of postinfection of four transcription factors. We sought to determine whether somatic genome reprogramming is completed by the onset of Oct4 reactivation. The complex regulation of the reactivation of inactive X chromosome (Xi) serves as a model for studying reprogramming of chromatin domains. A time-course analysis of the DNA methylation, gene expression, and X inactivation-specific transcript (Xist)/Tsix RNA fluorescence in situ hybridization revealed that expression of pluripotency- and tissue-specific marker genes was reset to the level of pluripotent stem cells within 2 days of fusion, whereas reprogramming of Xist/reactivation of Xi took at least 9 days. We found that trichostatin A, which normally activates gene expression, results in downregulation of Xist. This is due to activation of Dnmt3a and Tsix, two negative regulators of Xist. Moreover, delayed reprogramming of Xist/reactivation of inactive X chromosome after cell fusion was accelerated by DNA methylation and histone deacetylation of Xist, which follow upregulation of Dnmt3a and Tsix. STEM CELLS 2008;26:2821-2831
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