Recombinant CFTR detection in CF tracheal epithelial cells following in vitro liposome-mediated gene transfer

Alessia Colosimo, Stefania Scarpino, Federica Sangiuolo, Sabrina Di Sario, Giuseppe Mossa, Giuseppe Novelli, Bruno Dallapiccola

Research output: Contribution to journalArticlepeer-review

Abstract

The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-Chol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studying mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, while complete translocation of the recombinant CFTR to the apical membrane of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR processing and migration to the cell plasmalemma requires 72 h at least as demonstrated by immunocytofluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.

Original languageEnglish
Pages (from-to)723-729
Number of pages7
JournalBiochemistry and Molecular Biology International
Volume42
Issue number4
Publication statusPublished - Jul 1997

Keywords

  • Cationic liposomes
  • Cystic fibrosis
  • Gene therapy
  • Immunocytochemistry
  • Tracheal epithelial cells

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Molecular Biology

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