BACKGROUND: The cystic fibrosis transmembrane conductance regulator gene (CFTR) shows a complex pattern of expression. The regulatory elements conferring tissue-specific and temporal regulation are thought to lie mainly outside the promoter region. Previously, we identified DNase I hypersensitive sites (DHS) that may contain regulatory elements associated with the CFTR gene at -79.5 and at -20.5 kb with respect to the ATG and at 10 kb into the first intron. MATERIALS AND METHODS: In order to evaluate these regulatory elements in vivo we examined these DHS in a human CFTR gene that was introduced on a yeast artificial chromosome (YAC) into transgenic mice. The 310 kb human CFTR YAC was shown to restore the pheno-type of CF-null mice and so is likely to contain most of the regulatory elements required for tissue-specific expression of CFTR. RESULTS: We found that the YAC does not include the -79.5 kb region. The DHS at -20.5 kb is present in the chromatin of most tissues of the transgenic mice, supporting its non-tissue-specific nature. The DHS in the first intron is present in a more restricted set of tissues in the mice, although its presence does not show complete concordance with CFTR expression. The intron I DHS may be important for the higher levels of expression found in human pancreatic ducts and in lung submucosal glands. CONCLUSION: These data support the in vivo importance of these regulatory elements.
BACKGROUND: The cystic fibrosis transmembrane conductance regulator gene (CFTR) shows a complex pattern of expression. The regulatory elements conferring tissue-specific and temporal regulation are thought to lie mainly outside the promoter region. Previously, we identified DNase I hypersensitive sites (DHS) that may contain regulatory elements associated with the CFTR gene at -79.5 and at -20.5 kb with respect to the ATG and at 10 kb into the first intron. MATERIALS AND METHODS: In order to evaluate these regulatory elements in vivo we examined these DHS in a humanCFTR gene that was introduced on a yeast artificial chromosome (YAC) into transgenic mice. The 310 kb humanCFTR YAC was shown to restore the pheno-type of CF-null mice and so is likely to contain most of the regulatory elements required for tissue-specific expression of CFTR. RESULTS: We found that the YAC does not include the -79.5 kb region. The DHS at -20.5 kb is present in the chromatin of most tissues of the transgenic mice, supporting its non-tissue-specific nature. The DHS in the first intron is present in a more restricted set of tissues in the mice, although its presence does not show complete concordance with CFTR expression. The intron I DHS may be important for the higher levels of expression found in humanpancreatic ducts and in lung submucosal glands. CONCLUSION: These data support the in vivo importance of these regulatory elements.
Authors: S J Delaney; D P Rich; S A Thomson; M R Hargrave; P K Lovelock; M J Welsh; B J Wainwright Journal: Nat Genet Date: 1993-08 Impact factor: 38.330
Authors: J F Engelhardt; J R Yankaskas; S A Ernst; Y Yang; C R Marino; R C Boucher; J A Cohn; J M Wilson Journal: Nat Genet Date: 1992-11 Impact factor: 38.330
Authors: R E Ellsworth; D C Jamison; J W Touchman; S L Chissoe; V V Braden Maduro; G G Bouffard; N L Dietrich; S M Beckstrom-Sternberg; L M Iyer; L A Weintraub; M Cotton; L Courtney; J Edwards; R Maupin; P Ozersky; T Rohlfing; P Wohldmann; T Miner; K Kemp; J Kramer; I Korf; K Pepin; L Antonacci-Fulton; R S Fulton; P Minx; L W Hillier; R K Wilson; R H Waterston; W Miller; E D Green Journal: Proc Natl Acad Sci U S A Date: 2000-02-01 Impact factor: 11.205
Authors: Christopher J Ott; Neil P Blackledge; Jenny L Kerschner; Shih-Hsing Leir; Gregory E Crawford; Calvin U Cotton; Ann Harris Journal: Proc Natl Acad Sci U S A Date: 2009-11-06 Impact factor: 11.205