In vivo analysis of DNase I hypersensitive sites in the human CFTR gene

Danielle S. Moulin, Ania L. Manson, Hugh N. Nuthall, David J. Smith, Clare Huxley, Ann Harris*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

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 phenotype 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 1 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.

Original languageEnglish (US)
Pages (from-to)211-223
Number of pages13
JournalMolecular Medicine
Volume5
Issue number4
DOIs
StatePublished - 1999

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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