Gene targeting of CFTR DNA in CF epithelial cells.

摘要: A goal of cystic fibrosis (CF) gene therapy is correction the mutant CF transmembrane conductance regulator (CFTR) with wild-type (wt) DNA sequences to restore normal CFTR protein and function. Experiments wtCFTR cDNA expression vectors have shown that Cl ion transport phenotype associated can be corrected resemble in cells. An alternative cDNA-based strategies one corrects endogenous by targeted replacement wt homologue. To test whether such a strategy was feasible, small fragment homologous (SFHR) used replace specific genomic human epithelial Small fragments were transfected into transformed Replacement exogenous at appropriate locus its as mRNA indicated by: (1) allele-specific polymerase chain reaction (PCR) amplification mRNA-derived cDNA; (2) hybridization PCR products probes. In addition, functional activity determined whole cell patch clamp. Southern clamp analyses suggested approximately 1 100 cells underwent event resulted intact transport.