GENETIC STUDIES OF MOUSE CHROMOSOME 16 AND HUMAN 21

小鼠 16 号染色体和人类 21 号染色体的遗传学研究

• 批准号: 6241067
• 负责人: Roger H Reeves
• 金额: $ 20.24万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6241067
• 关键词: Downs syndrome; aneuploidy; animal genetic material tag; artificial chromosomes; cell fusion; chromosome 21; developmental genetics; embryonic stem cell; fungal genetics; gene dosage; genetic library; genetic mapping; genetic recombination; genetically modified animals; human genetic material tag; laboratory mouse; nucleic acid hybridization; polymerase chain reaction; transfection; transfection /expression vector; trisomy

The long term goal of Project II is to identify genes on mouse Chromosome 16 (Chr16) and human chromosome 21 (HC21) and to determine their contributions to maldevelopment when present at dosage imbalance. Functional analysis of large segments of these chromosomes will be effected by creating defined dosage imbalance in mice, using techniques developed through this program to isolate and modify yeast artificial chromosomes (YACs) and transfer them into murine embryonic stem (ES) cells. Modified ES cells will be analysed for integrity of the YAC- cloned segments. Appropriate lines will be used to make chimeras and thereby transmit YACs into the germ lines of mice, creating "YAC transgenic" animals with defined segmental aneuploidy. This approach will be used to refine mouse models to trisomy by creating dosage imbalance for genes on distal HC21 that are not represented in Chr16. The effects on development of genes in these segments will be assessed directly and after introduction into partial trisomy 16 mice (Project III). YAC transgenic mice will also be used to evaluate the effects of dosage imbalance for the conserved region of Chr16 and HC21 around the D21S55 locus. This region is associated with many characteristic of DS in "translocation DS" individuals who exhibit triplication of a subset of HC21 genes. This region is also likely to contain the homolog of the cerebellum observed in DS. Further YAC vector development will be pursued to establish a highly efficient method for identifying genes and mapping exons based on homologous recombination with specially constructed PCR-amplified cDNA libraries; to examine the application of YACs in homologous recombination-based approaches to gene deletion and the use of this approach to creation of segmental monosomy; and to characterize YAC integration sites in mammalian cells.

项目II的长期目标是鉴定小鼠染色体上的基因 16（CHR16）和人类染色体21（HC21），并确定其 出现在剂量不平衡的情况下，对马尔多发展的贡献。 这些染色体的大段的功能分析将是 通过使用技术在小鼠中产生定义的剂量不平衡而实现 通过该程序开发以隔离和修改酵母菌人工 染色体（YAC）并将其转移到鼠类胚胎茎（ES）中 细胞。 将分析修饰的ES细胞的YAC-的完整性 克隆段。 适当的线将用于制作嵌合体和 从而将YAC传输到小鼠的细菌中，创建“ YAC 转基因的“具有定义的节段性非整倍性的动物。这种方法 将通过产生剂量来将鼠标模型改进三体模型 远端HC21基因的不平衡，在CHR16中未表示。 将评估对这些段中基因发展的影响 直接并在引入部分三体三体16小鼠之后（项目 iii）。 YAC转基因小鼠也将用于评估 CHR16和HC21的保守区域的剂量不平衡周围 D21S55基因座。 该区域与DS的许多特征有关 在“易位DS”个人中表现出子集的三重三分之一 HC21基因。 该地区也可能包含 在DS中观察到小脑。 进一步的YAC向量发展将是 追求建立一种高效的方法来识别基因和 基于同源重组的映射外显子特别特别 构建的PCR扩增cDNA库；检查的应用 基于基于基因缺失和基于同源重组的方法中的YAC 使用这种方法来创建节段的单体手术；然后 表征哺乳动物细胞中YAC整合位点。