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Transcript analysis of CFTR nonsense mutations in lymphocytes and nasal epithelial cells from cystic fibrosis patients
Author(s) -
Will Katrin,
Dörk Thilo,
Stuhrmann Manfred,
Hardt Horst Von Der,
Ellemunter Helmut,
Tümmler Burkhard,
Schmidtke Jörg
Publication year - 1995
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380050305
Subject(s) - cystic fibrosis , biology , nonsense , nonsense mutation , nonsense mediated decay , cystic fibrosis transmembrane conductance regulator , genetics , gene , mutation , cancer research , immunology , microbiology and biotechnology , rna , rna splicing , missense mutation
The mutational effects at the mRNA level were investigated by RT‐PCR analysis of nine different nonsense mutations (Q39X, E60X, R75X, G542X, L719X, Y1092X, R1162X, S1196X, W1282X) and one frameshift mutation (1078delT) within the CFTR gene. With the exception of mutation R1162X, reduced mRNA levels ranging from 30% to less than 5% of the wild type have been observed. In case of the R75X and E60X mutations, the mRNA reduction was accompanied by the appearance of atypical CFTR isoforms. Single exon 3 skipping, as well as joint exon 2 and 3 skipping, was observed in lymphocyte and nasal epithelial mRNA derived from R75X alleles. The analysis of mRNA transcribed from E60X alleles revealed skipping of exon 3 (lymphocytes and nasal epithelial cells) or skipping of exons 3 and 4 (nasal epithelial cells). With the exception of the E60X mutation, no obvious tissue‐specific differences in the splicing pattern and ratios of mutation to wild‐type transcripts were detected between lymphocytes and nasal epithelial cells. In addition to aberrant splicing, the reduction of transcripts is the most common effect of nonsense and frameshift mutations within the CFTR gene. © 1995 Wiley‐Liss, Inc.