Proteolipidic vectors for gene transfer to the lung.
In order to develop improved synthetic gene transfer vectors, we have synthesized bifunctional peptides composed of a DNA binding peptide (P2) and ligand peptides selected by the phage display technique on tracheal epithelial cells. We have evaluated the capacity of these peptides to enhance the gene transfer efficiency of the cationic lipid DOTAP to the mouse lung. To optimize the in vivo transfection efficiency, we first compared the efficiency of DOTAP to transfect the lung by either intravenous injection or aerosolization. We then tested DNA/Peptide/DOTAP complexes formed at different Peptide/DNA and DOTAP/DNA charge ratios. Under optimal conditions, precompaction of DNA by peptide P2 gave a higher expression in the mouse lung using the luciferase reporter gene than DOTAP/DNA complexes. A further increase of transfection efficiency was obtained with the bifunctional peptide P2-9. Experiments performed with the GFP reporter gene showed expression in the alveolar parenchyme.
Vaysse
Laurence
L
Laboratoire de Pathologie Moléculaire et Thérapie Génique, Université Victor Segalen Bordeaux 2, 146 Rue Léo Saignat, Bordeaux Cedex, 33076, France.
Guillaume
Christine
C
Burgelin
Ingrid
I
Gorry
Philippe
P
Férec
Claude
C
Arveiler
Benoît
B
D000280:Administration, Inhalation; D000336:Aerosols; D000595:Amino Acid Sequence; D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D016503:Drug Delivery Systems; D005229:Fatty Acids, Monounsaturated / Q000008:administration & dosage / Q000493:pharmacokinetics; D005260:Female; D005456:Fluorescent Dyes / Q000008:administration & dosage / Q000493:pharmacokinetics; D005822:Genetic Vectors / Q000008:administration & dosage / Q000493:pharmacokinetics; D007275:Injections, Intravenous; D008024:Ligands; D008168:Lung / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D011510:Proteolipids / Q000008:administration & dosage* / Q000235:genetics* / Q000493:pharmacokinetics; D000644:Quaternary Ammonium Compounds / Q000008:administration & dosage / Q000493:pharmacokinetics; D014018:Tissue Distribution / Q000235:genetics; D014162:Transfection / Q000379:methods*
D000336:Aerosols; D004268:DNA-Binding Proteins; D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D008024:Ligands; D010446:Peptide Fragments; D011510:Proteolipids; D000644:Quaternary Ammonium Compounds; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane