Core C - The Mouse Intervention and Aging Core

项目总结(核心C) 鼠标干预和老龄化核心(核心C)将为方案项目(PPG)的调查人员提供 实现特定目标所需的小鼠模型，并将执行小鼠寿命测试、药物 治疗和鉴定小鼠的表型。这个PPG的实验集中在确定 逆转座子在衰老中的作用，确定控制反转录转座子和 关于测试反转座子干预。反转座子通过复制粘贴机制移动 造成DNA断裂、突变和基因组重排。此外，逆转录元件的转录 占基因组约45%的细菌可能会对宿主细胞造成损害。这三个人最近的研究 这个PPG的项目表明逆转录转座子在衰老过程中被激活。这一点的中心假设是 PPG是逆转录因子的激活有助于衰老的开始，而pRb和SIRT6蛋白 调节抑制性异染色质是负责抑制 线路-1(L1)。我们的初步数据表明，用逆转录酶抑制剂治疗SIRT6-/-小鼠， 抑制L1逆转座，缓解这些小鼠的早衰表型，为 我们的假设。核心C具体目标将是：(1)准备和维护机构动物护理和使用 本PPG内所有项目的委员会协议；(2)为项目调查人员饲养和维护小鼠；(3) 检测衰老对L1报告小鼠逆转座的影响；(4)检测过度活跃的L1的影响 对小鼠寿命和病理学的影响；(5)用于反转录转座子分析的pRb基因缺陷小鼠 激活；(6)用腺相关病毒(AAV)将Rb和SUV39H1基因导入老龄小鼠的肝脏；(7) 培育大脑特异性和成年发病的SIRT6基因敲除小鼠；(8)进行干预以抑制L1 NRTI在野生型和三种不同SIRT6缺陷小鼠模型中的逆转座及检测 反转录转座子活性和病理学；(9)确定NRTI是否延长了野生型和 构成全身SIRT6基因敲除小鼠；(10)产生和培育L1至L1-shRNA转基因小鼠 抑制L1转录。分析shRNA是否减轻野生型小鼠的增龄相关病理并延长 结构性SIRT6基因敲除小鼠的寿命和挽救病理；(11)SIRT6基因敲除小鼠的产生和繁殖 分离功能和调控位点突变以分析反转录转座子活性和能力 补充SIRT6基因敲除表型；(12)维护所有小鼠的数据库，以确保有效分布 向项目调查人员提供材料和数据。维护集中的啮齿动物群体将标准化 畜牧业条件、质量控制和生物样本用于整个PPG项目，改善 结果的重现性，并允许通过几种化验和项目对同一个体动物进行分析， 以及最大限度地减少动物使用。

PROJECT SUMMARY (CORE C) The Mouse Interventions and Aging Core (Core C) will provide investigators of the Program Project (PPG) with mouse models necessary to achieve their specific aims and will perform mouse lifespan assays, drug treatments and characterize mouse phenotypes. Experiments of this PPG focus on defining the role played by retrotransposable elements in aging, identifying the molecular mechanisms that control retrotransposons and on testing anti-transposon interventions. Retrotransposable elements move by a copy-and-paste mechanism creating DNA breaks, mutations and genomic rearrangements. Furthermore, the transcription of retroelements which comprise ~45% of the genome can take a toll on the host cell. The recent studies by all the three Projects of this PPG indicate that retrotransposons are activated during aging. The central hypothesis of this PPG is that the activation of retroelements contributes to the onset of aging and that pRb and SIRT6 proteins that regulate repressive heterochromatin are the key components of the pathways responsible for repression of LINE-1 (L1). Our preliminary data suggest that treating Sirt6-/- mice with reverse transcriptase inhibitors, which repress L1 retrotransposition, alleviates the premature aging phenotype of these mice, providing support for our hypothesis. Core C Specific Aims will be to: (1) prepare and maintain the Institutional Animal Care and Use Committee protocols for all projects within this PPG; (2) breed and maintain mice for project investigators; (3) examine the effect of aging on retrotransposition in L1 reporter mice; (4) examine the effect of hyperactive L1 on mouse lifespan and pathology; (5) breed pRb deficient mice for Project 1 for analysis of retrotransposon activation; (6) introduce Rb and Suv39h1 genes into livers of old mice using Adeno-Associated virus (AAV); (7) breed brain-specific and adult-onset Sirt6 knockout mice; (8) perform interventions to suppress L1 retrotransposition using nRTIs in wild type and three different Sirt6 deficient mouse models and examine retrotransposon activity and pathology; (9) determine whether nRTIs extend lifespan of wild type and constitutive whole body Sirt6 knockout mice; (10) generate and breed transgenic mice with shRNA to L1 to inhibit L1 transcription. Analyze whether shRNA alleviates age-related pathology of wild type mice and extends lifespan and rescues pathology of constitutive Sirt6 knockout mice; (11) generate and breed mice with Sirt6 separation of function and regulatory site mutations for analysis of retrotransposon activity and the ability to complement Sirt6 knockout phenotype; (12) maintain a database of all mice to ensure efficient distribution of materials and data to project investigators. Maintaining the centralized rodent colonies will standardize husbandry conditions, quality control and biological samples for use across the PPG projects, improve reproducibility of results, and allow the analysis of the same individual animals by several assays and projects, as well as minimize animal use.