TARGETED MUTAGENESIS OF WERNER SYNDROME GENE

维尔纳综合征基因的定向诱变

• 批准号: 6029831
• 负责人: JUNKO OSHIMA
• 金额: $ 22.62万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6029831
• 关键词: Werner's syndrome; aging; cytogenetics; developmental genetics; disease /disorder model; embryonic stem cell; gene expression; gene targeting; genetically modified animals; helicase; histopathology; laboratory mouse; model design /development; molecular cloning; pathologic process; prognosis; site directed mutagenesis

DESCRIPTION: The Werner syndrome (WS) ('Progeria of the Adult') is a rare autosomal recessive progeriod disorder. WS patients show a general appearance of premature aging, and exhibit premature onset of disorders commonly seen in the aged population including, bilateral ocular cataracts, type 2 diabetes mellitus, arteriosclerosis and osteoporosis. While they are susceptible to many neoplasms, these include a disproportionate number of cancers of mesenchymal origin and relatively rare neoplasms. Somatic cells from WS patients exhibit accelerated replicative senescence and a mutator phenotype. The Werner syndrome gene (WRN) has recently been shown to encode a helicase homologous to the RecQ of E. coli. Initially, four distinct mutations in the WRN gene were found in Japanese and Caucasian WS patients. Subsequently, more than a dozen different mutations have been identified. In this proposal, animal models of WS by targeted mutagenesis of the murine homolouge of WRN will be created. The goal is to create models of two naturally occurring human mutations, including the common Japanese exonic deletion. Life table parameters and anatomical pathology data, with emphasis upon analysis of the frequencies and the spectrum of neoplasms, will be obtained throughout the lifespan of C57B1/6 transgenic control mice. Primary cultures of somatic cells isolated from the transgenic mice will be used to determine replicative lifespan, WRN helicase activity levels and mutation frequencies at the HPRT locus. The long term goal is to elucidate the pathogenesis of WS and the role of WRN helicase in normal aging.

描述：Werner综合征（WS）（“成人的回忆”）是罕见的 常染色体隐性后足疾病。 WS患者显示一般 过早衰老的外观，并表现出疾病的过早发作 通常在老年人口中看到，包括双侧眼性白内障 2型糖尿病，动脉硬化和骨质疏松症。 当他们是 容易受到许多肿瘤的影响，其中包括不成比例的 间质起源和相对罕见的肿瘤的癌症。 体细胞 来自WS患者表现出加速的复制衰老和突变器 表型。 Werner综合征基因（WRN）最近已被证明编码一个解旋酶 与大肠杆菌的RECQ同源。 最初，四个不同的突变 在日本和高加索WS患者中发现了WRN基因。 随后，已经确定了十几个不同的突变。 在此提案中，通过针对鼠的靶向诱变的动物模型 将创建WRN的同性恋。 目标是创建两个模型 自然发生的人类突变，包括普通的日本外显子 删除。 生命表参数和解剖病理数据，带有 强调分析频率和肿瘤的光谱， 将在C57B1/6转基因对照小鼠的整个寿命中获得。 从转基因小鼠分离的体细胞的一级培养物将是 用于确定复制寿命，WRN解旋酶活性水平和 HPRT基因座的突变频率。 长期目标是阐明 WS的发病机理和WRN解旋酶在正常衰老中的作用。