A Murine Sox10 Dominant Negative COIN Allele for Functional Gene Analysis

描述（由申请人提供）：胶质细胞类型的胶质发生和维持对神经系统的发育和功能至关重要。Sox 10是一种发育转录因子，对多种神经胶质细胞谱系的发育至关重要，包括中枢神经系统中的少突胶质细胞以及神经嵴衍生的许旺细胞、卫星神经胶质细胞和肠神经系统神经元以及外周中的神经胶质细胞。在这些不同的人群中研究Sox10的功能一直很困难，因为简单的基因敲除会导致早期神经嵴祖细胞基因表达的完全丧失，从而导致胚胎死亡。暂时诱导Sox10丢失的努力受到mRNA和蛋白质衰变动力学的阻碍。在R03机制的背景下，我们提出在小鼠中产生Sox 10的C0N诱导（“COIN”）显性负等位基因作为分析基因功能的新工具。特异性目的1将产生在Sox10基因座中携带COIN盒的小鼠，其在Cre作用后导致荧光标记的显性阴性Sox10同种型的表达。具体目标2将定义Sox10COIN等位基因在COIN倒位前后对少突胶质细胞和肠神经嵴衍生谱系的影响。有条件地破坏Sox10在不同群体中的表达和功能的能力将通过为分析与祖细胞定向分化以治疗中枢和外周神经病相关的发育机制开辟途径来显著影响该领域。 公共卫生相关性：该提案的目标是开发一种新的小鼠突变体，可用于研究外周神经系统的发育和模型缺陷。了解神经系统如何发育和维持自身具有重要的临床意义，因为如果不这样做，可能会导致许多周围神经病变，从肌肉控制丧失到胃肠动力障碍，无法感知疼痛， 或温度变化和神经性疼痛。使用这样的小鼠模型有助于直接分化干细胞群体，用于治疗先天性疾病或恢复由于疾病或创伤引起的神经系统损伤。

DESCRIPTION (provided by applicant): Gliogenesis and maintenance of glial cell types are critical to development and function of the nervous system. Sox10 is a developmental transcription factor that is essential for development of multiple glial lineages including oligodendrocytes in the central nervous system as well as neural crest-derived Schwann cells, satellite glia and enteric nervous system neurons and glia in the periphery. Investigating Sox10 function in these distinct populations has been difficult because simple gene knockouts cause complete loss of gene expression in early neural crest progenitors resulting in embryonic lethality. Efforts to temporally induce loss of Sox10 have been hampered by kinetics of mRNA and protein decay. In the context of the R03 mechanism we propose generation of a COnditional INducible ("COIN") dominant negative allele of Sox10 in mice as a novel tool for analysis of gene function. Specific Aim 1 will generate mice bearing a COIN cassette in the Sox10 locus that upon Cre action results in expression of a fluorescently tagged dominant negative Sox10 isoform. Specific Aim 2 will define the effects of the Sox10COIN allele on oligodendrocyte and enteric neural crest-derived lineages before and after COIN inversion. The ability to conditionally disrupt Sox10 expression and function in distinct populations will significantly impact the field by opening avenues for analysis of developmental mechanisms that are relevant for directed differentiation of progenitors cells to treat central and peripheral neuropathies. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to develop a novel mouse mutant that can be used to investigate development and model deficits of the peripheral nervous system. Understanding how the nervous system develops and maintains itself has important clinical implications since failure to do so can result in a number of peripheral neuropathies ranging from loss of muscle control to gastrointestinal motility disorders, inability to sense pain or change in temperature and neuropathic pain. Using such mouse models helps direct differentiation of stem cell populations for treatment of congenital disorders or restoration of nervous system damage due to disease or trauma.