Recombinant CTFR detection in CF tracheal epithelial cells following in vitro liposomeme-mediated gene transfer.
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.
Cattedra di Genetica Umana e Medica, Dipartimento di Sanità Pubblica e Biologia Cellulare, Università "Tot Vergata" di Roma, Roma, Italy.
A
A
Colosimo
S
S
Scarpino
F
F
Sangiuolo
S D
SD
Sario
G
G
Mossa
G
G
Novelli
B
B
Dallapiccola