DEVELOPMENT OF CF MODEL AND CFTR ANALYSIS

囊性纤维化（CF）是最常见的常染色体隐性遗传疾病， 白种人 在人类CF患者中有缺陷的基因 最近被分离出来并鉴定了 该基因编码一种假定的 可能在控制离子转运中起作用的跨膜蛋白 穿过上皮细胞膜，因此被称为囊性 纤维化离子转运调节因子（CFTR）。 对人类的详细分析 CFTR基因揭示了一个单一的突变，这是目前在 约70%的CF患者。 到目前为止，所有接受检查的CF患者都有 该基因的一些修饰，表明CFTR中的缺陷是 导致CF的临床表现。 尚不清楚CFTR中的缺陷如何导致临床 在CF患者中观察到的症状。 本项目的目标是建立一个CF的动物模型， 用于研究这种疾病的基本缺陷，并检查 在正常和疾病状态下的CFTR。 详细分析基本 CF患者的缺陷是非常困难的，由于供应不足， 匹配正常和受影响的组织，并测试和评估新的 人类患者的治疗也很困难。 拟议研究 包括产生CF的小鼠模型，该模型在 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。 这种动物模型的制作 CF利用基因靶向（同源）的最新进展 重组）和小鼠胚胎干细胞的操作。 简言之， 将分离出小鼠CFTR基因的一部分， 类似于在许多人类CF患者中发现的， 植入老鼠的基因 这一过程的最终产品是生产 可用于建立具有一个 正常和一个突变CFTR基因;然后这些动物可以交配在一起 产生纯合的动物，这些动物有基因缺陷， 与大多数人类CF患者类似。 的可用性 合适的CF动物模型将使研究人员能够轻松地 检查缺陷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。 拟议 研究还将提供有关表达的详细信息， 正常小鼠的CFTR基因。

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.