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Recombinant CTFR detection in CF tracheal epithelial cells following in vitro liposomeme‐mediated gene transfer
Author(s) -
Colosimo Alessia,
Scarpino Stefania,
Sangiuolo Federica,
Sario Sabrina Di,
Mossa Giuseppe,
Novelli Giuseppe,
Dallapiccola Bruno
Publication year - 1997
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549700203151
Subject(s) - transfection , recombinant dna , microbiology and biotechnology , in vitro , cystic fibrosis , monoclonal antibody , messenger rna , liposome , biology , genetic enhancement , gene expression , chemistry , gene , antibody , biochemistry , immunology , genetics
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 immunocyto‐fluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.