DEVELOPMENT OF CF MODEL AND CFTR ANALYSIS

CF模型的建立和CFTR分析

• 批准号: 3245473
• 负责人: KEVIN A KELLEY
• 金额: $ 13.28万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3245473
• 关键词: cystic fibrosis; disease /disorder model; embryogenesis; embryonic stem cell; gene expression; gene mutation; genetic library; genetic manipulation; genetic recombination; genetically modified animals; human genetic material tag; in situ hybridization; laboratory mouse; microinjections; model design /development; northern blottings; nucleic acid probes; polymerase chain reaction; proteins; southern blotting; tissue /cell culture; tissue mosaicism; transfection

Cystic fibrosis (CF) is the most common autosomal recessive disorder of Caucasians. The gene which is defective in human CF patients has recently been isolated and characterized. This gene encodes a putative transmembrane protein that may play a role in controlling ion transport across epithelial membranes, and has consequently been termed the cystic fibrosis ion transport regulator (CFTR). Detailed analysis of the human CFTR gene has revealed a single mutation which is present in approximately 70% of CF patients. All CF patients examined to date have some modification of this gene, indicating that defects in CFTR are responsible for the clinical manifestations of CF. It is not known how the defects in CFTR result in the clinical manifestations observed in CF patients. The goal of this project is to create an animal model for CF which can be used to study the basic defect of this disease, and to examine the role of CFTR in the normal and disease state. Detailed analyses of the basic defect in CF patients is very difficult due to an inadequate supply of matched normal and affected tissue, and testing and evaluation of new treatments in human patients is also difficult. The proposed research involves producing a mouse model for CF which contains a mutation in the mouse CFTR gene that is analogous to the defect which has been detected in the majority of human CF patients. This production of an animal model for CF utilizes recent advances in gene targeting (homologous recombination) and manipulations of mouse embryonic stem cells. Briefly, a piece of the mouse CFTR gene will be isolated, and a mutation which is analogous to the one found in many human CF patients will be introduced into the mouse gene. The end product of this procedure is the production of mice which can be used to establish heterozygous lines having one normal and one mutant CFTR gene; these animals can then be mated together to produce homozygous animals that have the gene defect which is analogous to the majority of human CF patients. The availability Of an appropriate animal model for CF will allow investigators to easily examine the physiological effects of a defective CFTR gene, as well as providing a model for examining new treatment regimes. The proposed research will also provide detailed information concerning the expression of the normal mouse CFTR gene.

囊性纤维化是一种最常见的常染色体隐性遗传病。 高加索人。人类CF患者中存在缺陷的基因 最近被分离和鉴定。该基因编码一种推定的 可能在控制离子转运中发挥作用的跨膜蛋白 穿过上皮膜，因此被称为囊性 纤维化离子转运调节剂(CFTR)。对人类的详细分析 CFTR基因发现了一种单一突变，该突变存在于 大约70%的CF患者。到目前为止，所有接受检查的CF患者都有 对该基因的一些修饰，表明CFTR中的缺陷是 负责本病的临床表现。 目前尚不清楚CFTR的缺陷是如何导致临床的 在CF患者中观察到的表现。 该项目的目标是创建一种动物模型的CF，可以 用来研究本病的基本缺陷，并检验其作用 在正常和疾病状态下的CFTR值。详细分析了基本的 由于供应不足，CF患者的缺陷是非常困难的 匹配的正常组织和病变组织，以及新的 人类患者的治疗也很困难。拟议的研究 涉及到为CF制造一个小鼠模型，该模型包含一个突变的 与已检测到的缺陷相似的小鼠CFTR基因 在大多数人类CF患者中。这个动物模型的制作 For CF利用了基因靶向的最新进展(同源 重组)和小鼠胚胎干细胞的操作。简单地说， 将分离出一段小鼠CFTR基因，一个突变是 类似于在许多人类CF患者中发现的那种将被引入 转化成老鼠的基因。这一过程的最终产品是产品 可以用来建立杂合系的小鼠 正常和一个突变的CFTR基因；然后这些动物可以交配在一起 生产出具有基因缺陷的纯合子动物 类似于大多数人类CF患者。是否可以使用 合适的动物模型将使研究人员能够轻松地 检测有缺陷的cftr基因的生理效应，以及 为研究新的治疗方案提供了一个模型。建议数 研究还将提供有关该表达的详细信息 正常的小鼠CFTR基因。