TARGETED MUTAGENESIS OF WERNER SYNDROME GENE

维尔纳综合征基因的定向诱变

• 批准号: 2002353
• 负责人: JUNKO OSHIMA
• 金额: $ 18.88万
• 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/2002353
• 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：维尔纳综合征 (WS)（“成人早衰症”）是一种罕见的疾病 常染色体隐性早衰症。 WS 患者表现一般 出现过早衰老，并表现出疾病过早发作 常见于老年人群，包括双眼白内障、 2型糖尿病、动脉硬化和骨质疏松症。 虽然他们是 对许多肿瘤敏感，其中包括不成比例的数量 间质来源的癌症和相对罕见的肿瘤。 体细胞 WS 患者表现出加速的复制衰老和突变 表型。 沃纳综合征基因 (WRN) 最近被证明编码解旋酶 与大肠杆菌的 RecQ 同源。 最初，四种不同的突变 在日本和白人 WS 患者中发现了 WRN 基因。 随后，鉴定出十多种不同的突变。 在该提案中，通过小鼠定向诱变建立 WS 动物模型 将创建 WRN 的同源。 目标是创建两个模型 自然发生的人类突变，包括常见的日本外显子突变 删除。 生命表参数和解剖病理数据，具有 强调对肿瘤的频率和谱的分析， 将在 C57B1/6 转基因对照小鼠的整个生命周期中获得。 从转基因小鼠中分离的体细胞的原代培养物将被 用于确定复制寿命、WRN 解旋酶活性水平和 HPRT 位点的突变频率。 长期目标是阐明 WS 的发病机制以及 WRN 解旋酶在正常衰老中的作用。