Search of novel regulatory elements in the promoter region of CFTR gene

Over 1500 mutations have been identified in the CFTR gene so far, but the scanning of the whole coding region of the CFTR gene reveals about 92% of CF alleles. Thus, in about 10% of chromosomes from CF patients CFTR mutations are identified. Unknown mutations can be localized in non-coding regulatory CFTR regions. In particular, very few mutations have been found so far in the gene promoter. Expression of the CFTR gene shows a clear temporal and developmental regulation, but the regulatory mechanisms are poorly understood. Moreover, only few regulatory elements have been identified so far. Mutated promoter sequences might be unable to properly bind specific regulatory proteins or may give origin to new binding sites for different factors leading to the onset of the disease. Furthermore, the identification of novel protein factors involved in CFTR gene regulation in healthy individuals and in CF patients might result in a better understanding of molecular mechanisms underlying the disease and potentially, in the development of novel therapies. The aims of this project are: i) to analyze the promoter regions of CFTR gene to evaluate the presence of mutations in patients bearing “classic” and “atypical” CF (with one or both undetected mutations in coding regions of CFTR), compared with non CF control group; 2) to confirm the “causative” role of novel promoter mutations by “in vitro” studies of expression; 3) to search for novel regulatory elements that may interact with both normal and mutated promoters.