CONTROL OF N-CAM BY HOX AND PAX GENES IN TRANSGENIC MICE

HOX 和 PAX 基因对转基因小鼠中 N-CAM 的控制

• 批准号: 2403555
• 负责人: GERALD EDELMAN
• 金额: $ 28.57万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-15 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403555
• 关键词: aging; cell adhesion; cell adhesion molecules; cytogenetics; early embryonic stage; embryo /fetus tissue /cell culture; gene expression; gene mutation; genetic promoter element; genetically modified animals; genome; glycoproteins; insulin; membrane activity; microinjections; molecular cloning; neurofilament; neurons; regeneration; transposon /insertion element

Cell adhesion molecules (CAMs) play important roles in embryonic tissues and in maintaining the structure and function of adult tissues. Changes in CAM expression accompany a number of disease states, and it has been shown that they are involved in both regenerative and degenerative processes including nerve and muscle. We propose to study the function of CAMs in vivo by microinjecting into mouse embryos the gene for the chicken liver cell adhesion molecule L-CAM under the control of promoters that will cause it to be expressed in transgenic mice in locations where L-CAM is not normally seen. In addition we are making constructs to locations where L- CAM is not normally seen. In addition we are making constructs to truncate, mutate, and delete the endogenous gene for the neural cell adhesion molecule, N-CAM, by transfection of embryonicstem (ES) cells and transplantation into mouse blastocysts to produce transgenic mice. DNA constructs that include the chicken L-CAM gene coupled to the insulin promoter, the neurofilament promoter, and the crystallin promoter have been used in initial experiments in this laboratory to make transgenic mice. Those with the insulin promoter express chicken L-CAM in the beta-cells of the pancreas with some apparent perturbation of cell function. Cultures of ES cells have been established and mouse N-CAM genomic clones have been isolated. Production of mice ectopically expressing L-CAM under control of tissue specific promoters should provide opportunities to locally perturb function by L-CAM expression in the beta-cells of the pancreas, in neurons and in lens cells. These tissues do not express L-CAM normally but express other CAMs, can be assessed. Our preliminary studies show that some of these animals survive and can be used to look for the influence of L-CAM on regenerative processes (eg. nerve-muscle interactions) and degenerative processes that accompany aging. Alteration or ablation of the endogenous N-CAM gene should provide a detailed analysis of the role of each form of the molecule in development, regeneration, and degeneration in vivo. These studies should have extensive implications for a variety of birth defects and genetic defects, and provide important data for assessing the role of CAMs in events associated with aging.

细胞粘附分子（CAM）在胚胎组织中起重要作用 并维持成人组织的结构和功能。 变更 CAM表达伴随着许多疾病状态，并且已显示 他们参与再生和退化过程 包括神经和肌肉。 我们建议研究凸轮在 通过微注射到小鼠胚胎中的体内鸡肝基因 细胞粘附分子L-CAM在启动子的控制下会导致 它要在L-CAM不在的位置的转基因小鼠中表达 通常看到。 另外，我们正在建造构建到l-的位置 通常不会看到凸轮。 另外，我们正在建造 截断，突变并删除神经细胞的内源基因 通过转染胚胎（ES）细胞和粘附分子N-CAM和 移植到小鼠胚泡中以产生转基因小鼠。 DNA构建体，其中包括与胰岛素耦合的鸡L-CAM基因 启动子，神经丝启动子和结晶蛋白启动子已经是 在该实验室的初始实验中用于制造转基因小鼠。 那些胰岛素启动子在Beta细胞中的Express Chicken L-CAM 胰腺具有某种明显的细胞功能扰动。 文化 已经建立了ES细胞，小鼠N-CAM基因组克隆已经 孤立。 在控制下表达L-CAM的小鼠的产生 特定于组织的启动子应为本地扰动提供机会 L-CAM表达在胰腺的β细胞中的功能，在神经元中 在镜头细胞中。 这些组织不正常表达L-CAM，而是表达 可以评估其他凸轮。 我们的初步研究表明 这些动物生存，可用于寻找L-CAM对 再生过程（例如，神经肌肉相互作用）和退化性 伴随衰老的过程。 内源性的改变或消融 N-CAM基因应详细分析每种形式的作用 体内发育，再生和变性的分子。 这些 研究应该对各种出生缺陷具有广泛的影响 和遗传缺陷，并提供重要数据来评估 与衰老相关的事件中的凸轮。