KNOCKOUT MODEL FOR CANAVAN DISEASE

描述(摘自摘要)：天冬氨酸氨基转移酶(ASPA)和 N-乙酰天冬氨酸在Canavan病(CD)中的蓄积， 是1988年在我们实验室发现的。从那时起，我们已经诊断出 170多名Canavan病患者，其中三分之二是 德系犹太人血统。德系犹太人的携带率 我们实验室测定的个体是1/36，这表明 即使在非犹太人群体中，卡纳文病也更为常见， 比预想的要好。 纯化了天冬氨酸酰基酶，并对人、牛和小鼠进行了全长测定 已分离出ASPA基因，并定位了人ASPA基因 17号染色体的短臂。这项提议的目的是：1) 建立加那文病小鼠模型。小鼠的aspa基因已经被 特色化的。将在编码序列中引入中断，用于 创造基因敲除老鼠的目的。2)发展性和 将进行行为研究，以确定基因敲除小鼠的表型 型号将会确定。2)生化研究将测量尿液， 血液和脑NAA。脑部ASPA活性也将被测定。脑区 NAA也将在不牺牲动物的情况下在体内进行测定， 这对未来可能的治疗方法将是重要的。4) 将确定神经病理参数，试图确定 小鼠脑内出现的组织病理学变化。完全相同的 天冬氨酸氨基转移酶在脑中的细胞定位尚不清楚，尽管 它存在于白质中。转运蛋白基因的使用将使 用于ASPA的细胞定位。动物模型将是 在确定临床AM组织病理学时机方面的重要作用 鼠标的变化。5)未来将使用敲除鼠标 确定NAA在大脑中的代谢作用的研究 酶疗法和基因疗法的实验。

DESCRIPTION (from Abstract): The deficiency of aspartoacylase (ASPA) and the accumulation of N-acetylaspartic acid (NAA) in Canavan disease (CD), was discovered in our laboratory in 1988. Since then we have diagnosed more than 170 patients with Canavan disease, of whom 2/3 are of Ashkenazi Jewish extraction. The carrier rate among Ashkenazi Jewish individuals as determined in our laboratory is 1/36, suggesting that Canavan disease is much more common, even in non-Jewish populations, than expected. Aspartoacylase was purified, and full length human, bovine and mouse ASPA cDNA have been isolated and the human ASPA gene has been localized to the short arm of chromosome 17. The purpose of this proposal is: 1) Create a mouse model for Canavan disease. The mouse ASPA gene has been characterized. Disruption in the coding sequence will be introduced for the purpose of the creation of a knock-out mouse. 2) Developmental and behavioral studies will be done so the phenotype of the knock-out mouse model will be determined. 2) Biochemical studies will measure urinary, blood and brain NAA. Brain ASPA activity will also be determined. Brain NAA will also be determined in vivo without sacrificing the animals, which would be important for possible future therapies. 4) Neuropathological parameter will be determined, in an attempt to define the onset of histopathological changes in the mouse brain. The exact cellular localization of aspartoacylase in brain is unknown, although it is in the white matter. The use of the transporter gene will allow for the cellular localization of ASPA. The animal model will be important in establishing the timing of clinical AM histopathological changes in the mouse. 5) The knock-out mouse will be used in future studies for determining the metabolic role of NAA in brain, for experimentation with enzyme therapy and gene therapy.