Functional human CFTR produced by a stable minichromosome

Cristina Auriche, Daniela Carpani, Massimo Conese, Emanuela Caci, Olga Zegarra-Moran, Pierluigi Donini, Fiorentina Ascenzioni

Research output: Contribution to journalArticle


Artificial chromosomes have been claimed to be the ideal vector for gene therapy, but their use has been hampered by an inability to produce stable and well designed molecules. We have used a structurally defined minichromosome to clone the human cystic fybrosis transmembrane conductance regulator (CFTR) locus. To guarantee the presence of the proper regulatory elements, we used the 320 kb yeast artificial chromosome (YAC) 37AB12 with the intact CFTR gene and upstream sequences. The resulting minichromosome was analyzed for the presence of the entire CFTR gene and for its functional activity by molecular and functional methods. We have identified clones showing the presence of both the transcript and the CFTR protein. Moreover, in the same clones, a chloride secretory response to cAMP was detected. Mitotic and molecular stability after prolonged growth without selection demonstrated that the constructs were stable. This is the first example of a structurally known minichromosome made to contain an active therapeutic gene.

Original languageEnglish
Pages (from-to)862-868
Number of pages7
JournalEMBO Reports
Issue number9
Publication statusPublished - Sep 1 2002

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

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    Auriche, C., Carpani, D., Conese, M., Caci, E., Zegarra-Moran, O., Donini, P., & Ascenzioni, F. (2002). Functional human CFTR produced by a stable minichromosome. EMBO Reports, 3(9), 862-868.