PRODUCTION AND ANALYSIS OF APOE TRANSGENIC MICE

APOE转基因小鼠的制备和分析

• 批准号: 6218645
• 负责人: JOHN Ray GILBERT
• 金额: $ 22.94万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-15 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6218645
• 关键词: Alzheimer's disease; RNase protection assay; animal breeding; animal colony; animal genetic material tag; apolipoprotein E; cryopreservation; developmental genetics; egg /ovum; gene expression; genetic models; genetic transcription; genetically modified animals; immunoprecipitation; laboratory mouse; longitudinal animal study; model design /development; northern blottings; polymerase chain reaction; protein isoforms; southern blotting; western blottings

Alzheimer's disease *AD) is a devastating neurological disorder characterized by a progressive dementia which affects 5-10% of the elderly over age 65. Genetic factors have clearly been implicated in the early and late-onset familial forms of the linked disease. A form of late-onset AD has been show to be linked to the proximal long arm of chromosome 19 and subsequently associated with Apolipoprotein E (ApoE). Apo E is a plasma apolipoprotein that is involved in lipid transport and metabolism. Apo E is produced in brain and found in three major isoforms in the human population which have been designated E2. E3. E4. These alleles occur at a frequency of approximately 8, 76, and 16 per cent, respectively. Recently, an association between the ApoE4 allele and an increased risk of late-onset AD has been established. Subsequent studies have demonstrated that Apo E2 has a protective effect or delays the age of onset of Ad. The exact mechanism by which the different isoforms exert their effects are as yet unknown. To study the mechanisms by which Apo E exerts its effects and the role which Apo E plays in brain molecular biology, physiology, and metabolism awe are creating Apo E 2. 3. and 4 transgenic mice. Cosmid libraries from human AD patients with the appropriate Apo E genotype were prepared in the cosmid vectors pWE15 and SuperCos 1. Apo E containing cosmid clones were isolated and restriction mapped. Clones of the proper size containing the brain expression control regions for Apo E were produced and purified. Experiments to produce Apo E transgenic mice using microinjection procedures will be carried out using ice which lack a functional Apo E gene (Apo E "knockout" mice). Mice, lacking a functional murine Apo E, will be produced which are homozygous and heterozygous for the human Apo E2, 3 and 4 alleles. These mice will serve as a model system for the study of the role of Apo E in brain function and metabolism and its potential role in Alzheimer's disease.

阿尔茨海默病是一种破坏性的神经系统疾病 其特征在于影响5-10%的老年人的进行性痴呆 65岁以上 遗传因素显然与早期和 迟发性家族性连锁疾病 一种迟发性AD 已被证明与19号染色体的近端长臂有关， 随后与载脂蛋白E（ApoE）相关。 载脂蛋白E是一种血浆载脂蛋白，参与脂质转运， 新陈代谢. 载脂蛋白E在大脑中产生，并以三种主要亚型存在 在被命名为E2的人群中。 E3. E4. 这些 等位基因出现的频率约为8%、76%和16%， 分别 最近，ApoE 4等位基因与糖尿病之间的关联被发现。 迟发性AD的风险增加。 随后的研究 已经证明，载脂蛋白E2具有保护作用或延迟年龄， AD的开始 不同亚型发挥作用的确切机制 其影响尚不清楚。 探讨载脂蛋白E的作用机制， 载脂蛋白E在大脑分子生物学、生理学和新陈代谢中发挥作用 Ave正在制造载脂蛋白E 2。3. 4只转基因小鼠。 Cosmid库， 具有适当Apo E基因型的人AD患者在 粘粒载体pWE 15和SuperCos 1。 含Apo E的粘粒克隆 分离和限制性酶切图谱。 大小合适的克隆， 产生并纯化ApoE的脑表达控制区。 显微注射法制备载脂蛋白E转基因小鼠的实验 将使用缺乏功能性载脂蛋白E的冰块进行手术 基因（Apo E“敲除”小鼠）。 缺乏功能性鼠载脂蛋白E的小鼠， 将产生对于人Apo是纯合和杂合的 E2、3和4等位基因。 这些小鼠将作为研究的模型系统 载脂蛋白E在脑功能和代谢中的作用及其潜力 在阿尔茨海默病中的作用。