TY - JOUR
T1 - Enhanced efficiency of lactosylated poly-L-lysine-mediated gene transfer into cystic fibrosis airway epithelial cells
AU - Kollen, Wouter J.W.
AU - Schembri, Frank M.
AU - Gerwig, Gerrit J.
AU - Vliegenthart, Johannes F.G.
AU - Glick, Mary Catherine
AU - Scanlin, Thomas F.
PY - 1999
Y1 - 1999
N2 - Lactosylated poly-L-lysine is a nonviral vector that transfers genes into airway epithelial cells, including those from individuals with cystic fibrosis (CF). Substitution of 40% of the ε-amino groups of poly-L-lysine with lactosyl residues not only provided a ligand for receptor-mediated endocytosis, but also reduced the toxicity when compared with nonsubstituted poly-L-lysine. Lactosylated poly-L-lysine/pCMV-Luc complex is not toxic to cells in amounts that gave the maximum gene expression. The level of gene expression was regulated by using different combinations of chloroquine, glycerol, and E5CA peptide. Using cultured CF cells, chloroquine, combined with E5CA peptide, increased the transfer of the pCMVLucl lactosylated poly-L-lysine complex 10,000-fold compared with transfer without additives. In many systems, a high efficiency is of paramount importance and the enhancing agents can be used to modulate the expression of the gene. For example, transfer of pCMVLacZ/lactosylated poly-L-lysine complexes with chloroquine added to the transfection medium gave only 20% transfection efficiency of the reporter gene. However, when chloroquine was combined with glycerol, the efficiency was increased to 90%, thus approaching that reported with viral vectors. This highly efficient vector may be of great value for the future development of gene transfer systems.
AB - Lactosylated poly-L-lysine is a nonviral vector that transfers genes into airway epithelial cells, including those from individuals with cystic fibrosis (CF). Substitution of 40% of the ε-amino groups of poly-L-lysine with lactosyl residues not only provided a ligand for receptor-mediated endocytosis, but also reduced the toxicity when compared with nonsubstituted poly-L-lysine. Lactosylated poly-L-lysine/pCMV-Luc complex is not toxic to cells in amounts that gave the maximum gene expression. The level of gene expression was regulated by using different combinations of chloroquine, glycerol, and E5CA peptide. Using cultured CF cells, chloroquine, combined with E5CA peptide, increased the transfer of the pCMVLucl lactosylated poly-L-lysine complex 10,000-fold compared with transfer without additives. In many systems, a high efficiency is of paramount importance and the enhancing agents can be used to modulate the expression of the gene. For example, transfer of pCMVLacZ/lactosylated poly-L-lysine complexes with chloroquine added to the transfection medium gave only 20% transfection efficiency of the reporter gene. However, when chloroquine was combined with glycerol, the efficiency was increased to 90%, thus approaching that reported with viral vectors. This highly efficient vector may be of great value for the future development of gene transfer systems.
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U2 - 10.1165/ajrcmb.20.5.3417
DO - 10.1165/ajrcmb.20.5.3417
M3 - Article
C2 - 10226080
AN - SCOPUS:0033124961
VL - 20
SP - 1081
EP - 1086
JO - American Journal of Respiratory Cell and Molecular Biology
JF - American Journal of Respiratory Cell and Molecular Biology
SN - 1044-1549
IS - 5
ER -