Regulation and/or repression of cholinergic differentiation of murine embryonic stem cells using RNAi directed against transcription factor L3/Lhx8.
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.
Manabe
Takayuki
T
Department of 2nd Anatomy, Faculty of Medicine, Nara Medical University, Nara, Japan.
Wanaka
Akio
A
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D060850:LIM-Homeodomain Proteins; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology*; D014157:Transcription Factors
D018398:Homeodomain Proteins; C112960:LIM homeobox protein 8; D060850:LIM-Homeodomain Proteins; D014157:Transcription Factors