Abstract
Techniques for controlling the expression of a specific gene in embryonic stem cells are effective and important for clarifying the functions of the gene. Regarding differentiation of cells into nervous system components, these techniques would play key roles in elucidating, not only the differentiation mechanisms of neuronal and glial cells, but also how neuronal phenotypes are determined. In this chapter, we describe an RNA interference method for suppressing cholinergic differentiation in murine embryonic stem cells by knockdown of expression of the transcription factor L3/Lhx8, a Lim homeobox gene family protein. This method will greatly facilitate functional analyses of the factors involved in neuronal differentiation and regeneration and contribute to cell transplantation studies.
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Acknowledgments
We thank Dr. H. Niwa (RIKEN) for providing the murine ES cells. T.M. was supported by a grant from The Ichiro Kanehara Foundation and a Grant-in-Aid for Scientific Research on Priority Areas, Advanced Brain Science Project, from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
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Manabe, T., Wanaka, A. (2010). Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8. In: Zhang, B., Stellwag, E. (eds) RNAi and microRNA-Mediated Gene Regulation in Stem Cells. Methods in Molecular Biology, vol 650. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-769-3_8
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DOI: https://doi.org/10.1007/978-1-60761-769-3_8
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