CORE--MOLECULAR AND CELLULAR ASSAY FACILITY

（申请人描述）核心A，分子和细胞分析的长期目标是[1]提供WRN野生型、杂合型和非人类和小鼠来源的细胞、细胞系或组织的基因型和表型特征，[2]为项目获取和储存肉瘤和脑膜瘤，以便进行WRN序列分析。这些目标将通过以下具体目标来实现：在通过等位基因转移操纵WRN表达的人类细胞系中；从敲除小鼠的组织中提取的细胞和细胞系；在酵母中，Core将提供：a.通过细胞遗传学检查对基因组不稳定性进行分析，以确定维尔纳综合征的总体染色体重排特征，包括整个染色体的丢失或获得以及杂色易位嵌合体。b.在RNA水平上评估WRN的表达。c.验证WRN基因突变的DNA序列分析。2. 对于手术切除获得的肉瘤和脑膜瘤，Core将收集和储存组织，用于组织学表征和DNA和RNA分离，用于项目项目，并对配对的正常和肿瘤样本进行WRN DNA序列分析。

DESCRIPTION: (Applicant's Description) The long term goals of Core A, Molecular and Cellular Assays, are [1] to provide genotypic and phenotypic characterization of WRN wild-type, heterozygous, and null human- and mouse-derived cells, cell lines, or tissues generated by this program, and [2] to procure and bank sarcomas and meningiomas for the Program Project in order to perform WRN sequence analysis. These goals will be achieved by the following specific aims: 1. In human cell lines in which WRN expression has been manipulated by allele transfer; in cells and cell lines derived from tissues of knock out mice; and in yeast the Core will provide: a. analysis of genomic instability by cytogenetic examination for gross chromosomal rearrangements characteristic of Werner's syndrome, including whole chromosome loss or gain and variegated translocation mosaicism. b. assessment of WRN expression at the RNA level. c. DNA sequence analysis for verification of WRN gene mutations. 2. For sarcomas and meningiomas obtained from surgical resections, the Core will collect and bank tissue for histologic characterization and DNA and RNA isolation for use in the Program Project and perform WRN DNA sequence analysis of paired normal and tumor samples.