DEVELOPMENT OF CF MODEL AND CFTR ANALYSIS

CF模型的建立和CFTR分析

• 批准号: 3245471
• 负责人: KEVIN A KELLEY
• 金额: $ 12.35万
• 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/3245471
• 关键词: cystic fibrosis; disease /disorder model; embryogenesis; 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（CF）是最常见的常染色体隐性遗传病 白种人。 人类 CF 患者中存在缺陷的基因 最近被分离和表征。 该基因编码一个假定的 可能在控制离子运输中发挥作用的跨膜蛋白 穿过上皮细胞膜，因此被称为囊性 纤维化离子转运调节剂（CFTR）。 人体细节分析 CFTR 基因揭示了存在于 大约 70% 的 CF 患者。 迄今为止接受检查的所有 CF 患者均患有 该基因的一些修饰，表明 CFTR 存在缺陷 是 CF 临床表现的原因。 目前尚不清楚 CFTR 的缺陷如何导致临床结果 CF 患者中观察到的表现。 该项目的目标是创建一个 CF 动物模型 用于研究本病的基本缺陷，并检验其作用 正常和疾病状态下的CFTR。 详细分析基本情况 由于供应不足，CF患者的缺陷非常困难 匹配正常和受影响的组织，并测试和评估新的 对人类患者的治疗也很困难。 拟议的研究 涉及制作 CF 小鼠模型，其中包含 与已检测到的缺陷类似的小鼠 CFTR 基因 在大多数人类 CF 患者中。 本次动物模型的制作 CF 利用基因靶向的最新进展（同源 重组）和小鼠胚胎干细胞的操作。 简要地， 将分离出小鼠 CFTR 基因的一部分，并进行突变 将介绍与许多人类 CF 患者中发现的类似物 进入小鼠基因。 此过程的最终产品是生产 的小鼠，其可用于建立具有一个的杂合系 正常和一个突变的 CFTR 基因；然后这些动物可以交配在一起 产生具有基因缺陷的纯合动物 类似于大多数人类 CF 患者。 的可用性 合适的 CF 动物模型将使研究人员能够轻松 检查有缺陷的 CFTR 基因的生理影响，以及 为检查新的治疗方案提供模型。 拟议的 研究还将提供有关表达的详细信息 正常小鼠 CFTR 基因。