Manabe Takayuki, Wanaka Akio
Department of Second Anatomy, Faculty of Medicine, Nara Medical University, 840 Shijo-cho, Kashihara, Nara, 634-8521, Japan.
Department of Neuroanatomy and Neuropharmacology, Faculty of Nursing, Chukyogakuin University, 2216 Toki-cho, Mizunami, Gifu, 509-6192, Japan.
Methods Mol Biol. 2017;1622:101-109. doi: 10.1007/978-1-4939-7108-4_8.
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 a 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 will contribute to cell transplantation studies.
控制胚胎干细胞中特定基因表达的技术对于阐明该基因的功能是有效且重要的。关于细胞分化为神经系统成分，这些技术不仅在阐明神经元和神经胶质细胞的分化机制方面，而且在确定神经元表型的方式方面都将发挥关键作用。在本章中，我们描述了一种RNA干扰方法，通过敲低转录因子L3/Lhx8（一种Lim同源框基因家族蛋白）的表达来抑制小鼠胚胎干细胞中的胆碱能分化。该方法将极大地促进对参与神经元分化和再生的因子的功能分析，并将有助于细胞移植研究。
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