Stem Cells http://www.stemcellsportal.com/
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

First published online February 2, 2006
OPEN ACCESS ARTICLE
This Article
Free via Open Access: OA
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrowOA All Versions of this Article:
2005-0475v1
24/6/1467    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Reprints/Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hough, S. R.
Right arrow Articles by Wiederholt, K. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hough, S. R.
Right arrow Articles by Wiederholt, K. A.
Submitted on September 28, 2005
Accepted on January 22, 2006

Embryonic Stem Cells

Differentiation of mouse embryonic stem cells following RNAi-mediated silencing of OCT4 and Nanog

Shelley R. Hough 1*, Ian Clements 2, Peter J. Welch 1, Kristin A. Wiederholt 1

1 Research and Development, Gene Regulation Group, Invitrogen Corporation, Carlsbad, California
2 Molecular Probes, Eugene, Oregon

* To whom correspondence should be addressed. E-mail: shelley.hough{at}invitrogen.com.


  Abstract

RNAi holds great promise as a tool to study the basic biology of stem cells or to direct differentiation in a specific manner. Barriers to achieving efficient and specific gene silencing in RNAi experiments include limitations in transfection efficiency and in the efficacy and specificity of RNAi silencing effectors. Here, we combine methods of efficient lipid-mediated delivery with chemically modified RNAi compounds to silence genes related to pluripotency, in order to direct differentiation of mouse embryonic stem cells. Following transfection of embryonic stem cells with OCT4- or Nanog-targeted RNAi compounds, levels of OCT4 or Nanog transcript and protein were reduced accordingly. Reduction in OCT4 expression correlated with induction of trophectoderm genes Cdx2, Hand1 and PL-1, with formation of cells with trophoblast giant cell phenotype after 6 days. Reduction in Nanog expression correlated with induction of extra-embryonic endoderm genes GATA4, GATA6 and laminin B1, with subsequent generation of groups of cells with parietal endoderm phenotype. Our results indicate that transient inhibition of OCT4 or Nanog by RNAi compounds is sufficient to induce differentiation towards extraembryonic lineages, which supports the model that these transcription factors function in a dose-dependent manner to influence cell fate.

Key Words. OCT4, Nanog, mouse embryonic stem cells, RNAi, trophectoderm, primitive endoderm




This article has been cited by other articles:


Home page
 Stem CellsHome page
S. He, D. Pant, A. Schiffmacher, A. Meece, and C. L. Keefer
Lymphoid Enhancer Factor 1-Mediated Wnt Signaling Promotes the Initiation of Trophoblast Lineage Differentiation in Mouse Embryonic Stem Cells
Stem Cells, April 1, 2008; 26(4): 842 - 849.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. Wang, T. W. Theunissen, and S. H. Orkin
Site-directed, virus-free, and inducible RNAi in embryonic stem cells
PNAS, December 26, 2007; 104(52): 20850 - 20855.
[Abstract] [Full Text] [PDF]

Home page
 J EndocrinolHome page
M. Zhao, S. A Amiel, M. R Christie, P. Muiesan, P. Srinivasan, W. Littlejohn, M. Rela, M. Arno, N. Heaton, and G. C. Huang
Evidence for the presence of stem cell-like progenitor cells in human adult pancreas
J. Endocrinol., December 1, 2007; 195(3): 407 - 414.
[Abstract] [Full Text] [PDF]

Home page
 Stem CellsHome page
M. Choudhary, X. Zhang, P. Stojkovic, L. Hyslop, G. Anyfantis, M. Herbert, A. P. Murdoch, M. Stojkovic, and M. Lako
Putative Role of Hyaluronan and Its Related Genes, HAS2 and RHAMM, in Human Early Preimplantation Embryogenesis and Embryonic Stem Cell Characterization
Stem Cells, December 1, 2007; 25(12): 3045 - 3057.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
L. S. Lim, Y.-H. Loh, W. Zhang, Y. Li, X. Chen, Y. Wang, M. Bakre, H.-H. Ng, and L. W. Stanton
Zic3 Is Required for Maintenance of Pluripotency in Embryonic Stem Cells
Mol. Biol. Cell, April 1, 2007; 18(4): 1348 - 1358.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
B. Boer, J. Kopp, S. Mallanna, M. Desler, H. Chakravarthy, P. J. Wilder, C. Bernadt, and A. Rizzino
Elevating the levels of Sox2 in embryonal carcinoma cells and embryonic stem cells inhibits the expression of Sox2:Oct-3/4 target genes
Nucleic Acids Res., March 27, 2007; (2007) gkm059v2.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Qin, T. Yu, T. Qing, Y. Liu, Y. Zhao, J. Cai, J. Li, Z. Song, X. Qu, P. Zhou, et al.
Regulation of Apoptosis and Differentiation by p53 in Human Embryonic Stem Cells
J. Biol. Chem., February 23, 2007; 282(8): 5842 - 5852.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
L. Pereira, F. Yi, and B. J. Merrill
Repression of Nanog Gene Transcription by Tcf3 Limits Embryonic Stem Cell Self-Renewal
Mol. Cell. Biol., October 15, 2006; 26(20): 7479 - 7491.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
STEM CELLS THE ONCOLOGIST CME ALPHAMED PRESS JOURNALS

Copyright © 2006 by AlphaMed Press.