A Murine Sox10 Dominant Negative COIN Allele for Functional Gene Analysis

描述(由申请人提供)：胶质细胞类型的形成和维持对神经系统的发育和功能至关重要。SOX10是一种发育中的转录因子，对多种神经胶质细胞的发育至关重要，包括中枢神经系统中的少突胶质细胞以及周围神经脊源性雪旺细胞、卫星胶质细胞和肠神经系统神经元和神经胶质细胞。在这些不同的种群中研究Sox10的功能一直是困难的，因为简单的基因敲除会导致早期神经脊祖细胞基因表达的完全丧失，从而导致胚胎死亡。导致Sox10基因暂时丢失的努力受到了mRNA和蛋白质衰变动力学的阻碍。在R03机制的背景下，我们建议在小鼠中产生一个条件诱导的Sox10的显性负等位基因，作为一种新的基因功能分析工具。特定目的1将产生在Sox10基因座上携带硬币盒的小鼠，Cre作用后会导致荧光标记的显性负性Sox10亚型的表达。具体目标2将确定Sox10COIN等位基因在硬币倒置前后对少突胶质细胞和肠神经脊源性谱系的影响。在不同人群中条件性干扰Sox10表达和功能的能力将为分析与祖细胞定向分化相关的发育机制开辟道路，从而治疗中枢和外周神经病。

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.