Reverse genetics and cystic fibrosis.
Reverse genetics and cystic fibrosis.
复制标题
反向遗传学和囊性纤维化。
DOI：
10.1165/ajrcmb/2.4.309
复制
发表时间：
1990
影响因子：
6.4
通讯作者：
Collins,FS
中科院分区：
文献类型：
--
作者：
Iannuzzi,MC;Collins,FS
The protein responsible for cystic fibrosis has been identified using an approach called" reverse" genetics. This approach relies on the chromosomal map position to direct the search for a disease gene, several novel cloning strategies to isolate the gene, and the gene's sequence to define the abnormal protein. Reverse genetics, because it does not require prior knowledge of the protein's biochemical function, has wide utility and is being used to define the defects in many single-gene disorders. This update presents the reverse genetics approach and uses cystic fibrosis to illustrate the principles involved.The standard biochemical approach to the molecular basis of inherited disorders has often been hampered by a lack of functional assays, a low abundance of abnormal gene products, the complexity of cellular and protein interactions, and even the inability to identify the affected cell type. An alternative approach without regard to cellular pathophysiology or biochemical data has recently been developed. This approach is based on locating (mapping) the disease gene in the human genome and using the map position to clone, sequence, and determine the abnormal gene product. This process has been called" reverse" genetics, because it is the reverse of the usual approach of isolating the protein and then using the protein to clone the gene (1). In collaboration with investigators at the Hospital for Sick Children in Toronto, we recently used reverse genetics to identify the abnormal protein responsible for cystic fibrosis (CF), the cystic fibrosis transmembrane regulator (CFTR)(2-4). Although biochemical and electrophysiologic investigations previously detected a physiologic abnormality in CF-an abnormal ion transport in the apical membrane of epithelial cells-the CF protein was not forthcoming. By mapping the gene to the long arm of chromosome 7 and then honing in on and cloning the CF gene, the responsible protein was identified.
