KNOCKOUT MODEL FOR CANAVAN DISEASE

描述（来自摘要）：β-淀粉酰化酶（ASPA）和 N-乙酰天冬氨酸（NAA）在卡纳万病（CD）中积累， 是1988年在我们实验室发现的从那以后，我们诊断出 超过170例Canavan病患者，其中2/3为 德系犹太血统。德系犹太人的携带率 在我们的实验室中确定的个体是1/36，这表明， 卡纳万病更常见，即使在非犹太人中， 比预期 纯化了天冬酰胺酰化酶，并制备了全长人、牛和小鼠天冬酰胺酰化酶 已分离出ASPA cDNA，并已定位人ASPA基因 17号染色体的短臂上本提案的目的是：1） 建立卡纳万病的小鼠模型。小鼠ASPA基因已经被 表征了将引入编码序列的中断， 创造一只基因敲除小鼠的目的。2)发展和 将进行行为研究，以便敲除小鼠的表型 模型将被确定。2)生化研究将测量尿液， 血液和脑NAA。还将测定脑ASPA活性。大脑 还将在不处死动物的情况下在体内测定NAA， 这对未来可能的治疗很重要。四、 将确定神经病理学参数，以尝试定义 小鼠脑组织病理学变化的开始。的确切 β-乙酰化酶在脑中的细胞定位尚不清楚，尽管 它在白色物质中。使用这种转运基因 用于ASPA的细胞定位。动物模型将是 重要的是建立临床AM组织病理学的时间 鼠标的变化5)敲除小鼠将在未来使用 研究确定NAA在大脑中的代谢作用， 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.