TARGETED MUTAGENESIS OF WERNER SYNDROME GENE

描述：Werner综合征(WS)(成人早衰症)是一种罕见的疾病 常染色体隐性遗传性早孕症。WS患者表现出一般情况 出现过早衰老，并表现出过早发病的障碍 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。当他们在 容易患上许多肿瘤，其中包括数量不成比例的 起源于间充质组织的癌症和相对罕见的肿瘤。体细胞 来自WS患者的复制衰老加速和突变子 表型。 沃纳综合征基因(WRN)最近被证明编码一种解旋酶 与大肠杆菌的RecQ同源。最初，四种截然不同的突变 在日本和高加索WS患者中发现了WRN基因。 随后，发现了十几种不同的突变。 在这项建议中，通过对小鼠进行靶向突变建立WS的动物模型 将创建WRN的同色。目标是创建两个人的模型 自然发生的人类突变，包括常见的日语外显子 删除。生命表参数和解剖病理数据， 重点分析肿瘤的频率和频谱， 将在C57B1/6转基因对照小鼠的整个生命周期内获得。 从转基因小鼠分离的体细胞的原代培养将是 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。我们的长期目标是阐明 WS的发病机制及WRN解旋酶在正常衰老中的作用

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.