Spatio-temporal control of FOXO3

描述（由申请人提供）：在C工作。elegans已经确定了一种控制寿命和对压力反应的信号通路。胰岛素-生长因子受体（Daf-2）或PI 3 'K（年龄-1）的功能等位基因的丧失导致2- 3倍的寿命延长和对应激源如UV照射、氧化损伤和热的强大抵抗力。所有这些有益的作用都是由转录因子Daf-16介导的。虽然Daf-16广泛表达，但促长寿效应是由特定组织中的Daf-16活性介导的。这表明Daf-16参与组织之间的对话，确保对环境变化和代谢功能的协调反应。哺乳动物有四个Daf-16直系同源物，FOXO 3是遗传学上最接近的-基于序列和功能。促进FOXO 3的核定位在多种神经变性模型中具有强烈的神经保护作用。了解FOXO 3在体内的作用将需要时空控制其核定位。在这 我们将制造转基因小鼠，其中FOXO 3融合到雌激素受体的配体结合结构域（一种称为“ERT”的他莫昔芬敏感型）。当插入到CAGGS-floxed-STOP载体中时，我们将能够以细胞类型特异性方式表达FOXO 3，并通过给予他莫昔芬驱动其核。在这个提议中，我们将表征这些小鼠的转基因泄漏，组织特异性表达，脱靶效应和FOXO 3靶基因表达的他莫昔芬诱导。该提案的交付是一种多功能小鼠，允许FOXO 3基因转录的时空控制。 公共卫生相关性：在模型生物和培养皿中进行的观察表明，一种名为FOXO 3的特定蛋白质对引起肌萎缩性侧索硬化症（ALS）（也称为Lou Gehrig病）的损伤具有深刻的保护作用。在这个“工具开发”的建议中，我将制作一只小鼠，使我们能够研究FOXO 3在小鼠疾病模型中的时空作用。

DESCRIPTION (provided by applicant): Work in C. elegans has defined a signaling pathway that controls longevity and response to stress. Loss of function alleles of the insulin-growth factor receptor (Daf-2) or PI3'K (age-1) lead to 2-3x longer life and powerful resistance to stressors such as UV irradiation, oxidative insult and heat. All of these beneficial actions are mediated by the transcription factor, Daf-16. While Daf-16 is widely expressed, the pro-longevity effects are mediated by Daf-16 activity in specific tissues. This indicates that Daf-16 participate in a conversation among tissues that ensures coordinated responses to environmental changes and metabolic function. Mammals have four Daf-16 orthologs and FOXO3 is the phylogenetically the closest - based on sequence and function. Promotion of nuclear localization of FOXO3 is strongly neuroprotective in multiple models of neurodegeneration. Understanding the role of FOXO3 in vivo will require spatio-temporal control of its nuclear localization. In this tool-development proposal we will make transgenic mice in which FOXO3 is fused to the ligand binding domain of the estrogen receptor (a tamoxifen-sensitive version called "ERT"). When inserted into the CAGGS-floxed-STOP vector we will be able to express FOXO3 in a cell-type specific manner and drive it nuclear by administration of tamoxifen. In this proposal we will characterize these mice for transgene leakiness, tissue specific expression, off target effects and tamoxifen inducibility of FOXO3 target gene expression. The deliverable of this proposal is a versatile mouse that permits spatio-temporal control of FOXO3 gene transcription. PUBLIC HEALTH RELEVANCE: Observations made in model organisms and petri dishes indicate that a specific protein called FOXO3 has profound protective properties against insults that cause Amyotrophic Lateral Sclerosis (ALS), also known as Lou Gehrig's Disease. In this "tool- development" proposal I will make a mouse that will allow us to study the spatio- temporal role of FOXO3 in mouse models of disease.