MOLECULAR STUDIES OF GENOMIC IMPRINTING IN HUMANS

人类基因组印记的分子研究

• 批准号: 6520933
• 负责人: Robert D Nicholls
• 金额: $ 31.8万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520933
• 关键词: Prader Willi syndrome; clinical research; gene deletion mutation; gene expression; gene mutation; genetic disorder; genetic polymorphism; genetic regulation; genetic screening; genetically modified animals; genomic imprinting; happy puppet syndrome; human subject; laboratory mouse; molecular cloning; molecular genetics; nucleic acid sequence; phenotype; polymerase chain reaction; yeast two hybrid system

DESCRIPTION: (Adapted from investigator's abstract) This is a revised application for a competitive renewal of a grant to study the molecular mechanisms of imprinting disorders. Prader-Willi syndrome (PWS) is a devastating, complex developmental and neurobehavioral disorder which results from loss of paternal imprinted gene expression at chromosome 15q11-q13. The investigator and others have defined a series of paternally expressed imprinted genes in this chromosome region and the syntenic mouse region in chromosome 7C. The investigator has recently developed a novel mouse model for PWS, and Angelman syndrome (AS), in which a transgene insertion has deleted all the PWS/AS homologous genes. A severe neonatal failure-to-thrive, typical of PWS in humans, occurs on paternal inheritance, but the transgenic line is maintained by maternal transmission. This PWS mouse model will be used to determine which genes play which roles in PWS, and to define the biochemical basis of the disorder. The main hypotheses are that PWS is a contiguous gene syndrome with multiple imprinted genes contributing to the PWS phenotype, and that these genes can be uniquely identified by transgenic rescue of the PWS mouse model. The goal of this proposal is to genetically dissect the imprinted gene(s) underlying each component of the PWS and PWS mouse model phenotypes by an approach consisting of three Specific Aims: (1) use of transgenic rescue of the PWS-associated phenotypes in the PWS mouse model, in order to define specific genes for each phenotypic component. Transgenic technology utilizing genomic clones of varying gene content from the 2 Mb PWS region, and cDNA clones, will be done to rescue the PWS-related phenotypes; (2) to characterize the function of genes shown to be responsible for phenotypic aspects of PWS; and (3) to screen PWS or PWS-like patients with no known 15q11-q13 molecular abnormality, or those patients with single phenotypic components of PWS, for mutations in each candidate gene for specific phenotypic components identified by mouse models.

描述：（改编自研究者的摘要）这是一项修订后的申请，旨在竞争性更新拨款，以研究印记疾病的分子机制。普瑞德威利综合征 (PWS) 是一种破坏性、复杂的发育和神经行为障碍，由染色体 15q11-q13 父系印记基因表达缺失所致。研究人员和其他人在该染色体区域和 7C 号染色体的同线小鼠区域中定义了一系列父系表达的印记基因。研究人员最近开发了一种新的 PWS 和天使综合征 (AS) 小鼠模型，其中转基因插入删除了所有 PWS/AS 同源基因。严重的新生儿发育不良是人类 PWS 的典型现象，发生在父系遗传中，但转基因系是通过母系传播维持的。该 PWS 小鼠模型将用于确定哪些基因在 PWS 中发挥哪些作用，并确定该疾病的生化基础。主要假设是，PWS 是一种连续基因综合征，具有多个导致 PWS 表型的印记基因，并且这些基因可以通过 PWS 小鼠模型的转基因拯救来唯一识别。本提案的目标是通过由三个具体目标组成的方法，从基因角度剖析 PWS 和 PWS 小鼠模型表型每个组成部分背后的印记基因：（1）在 PWS 小鼠模型中使用转基因拯救 PWS 相关表型，以便为每个表型组成部分定义特定基因。转基因技术将利用 2 Mb PWS 区域不同基因含量的基因组克隆和 cDNA 克隆来挽救 PWS 相关表型； (2) 表征与 PWS 表型相关的基因的功能； (3) 筛查没有已知 15q11-q13 分子异常的 PWS 或 PWS 样患者，或具有 PWS 单一表型成分的患者，针对小鼠模型鉴定的特定表型成分的每个候选基因中的突变。