Variable levels of normal RNA in different fetal organs carrying a cystic fibrosis transmembrane conductance regulator splicing mutation

Disease severity varies among cystic fibrosis (CF) patients carrying the sa me cystic fibrosis transmembrane conductance regulator (CFTR) genotype and among organs of the same individual. It has been shown that the class V spl icing mutation 3849 + 10 kb C-->T produces both normal and aberrantly splic ed CFTR transcripts. We analyzed the levels of normal CFTR messenger RNA (m RNA) in different organs of an aborted fetus carrying the 3849 + 10 kb C--> T mutation, and found that they correlated with the histopathologic changes observed in these organs. We performed semiquantitative nondifferential re verse transcription-polymerase chain reaction on several organs from a 22-w k aborted CF fetus carrying the 3849 + 10 kb C-->T mutation. A very low lev el (1%) of normal CFTR mRNA was detected in the severely affected ileum of this fetus. Higher levels were found in the histopathologically unaffected trachea (17%), colon (19%), and lung (26%). Thus, as early as in utero, the regulation of alternative splice-site selection is an important mechanism underlying variable CF severity. Understanding of the mechanisms regulating alternative splicing in different tissues will contribute to potential the rapy for patients carrying splicing mutations in CF and other human disease genes.