Intratracheal gene delivery to the mouse airway: characterization of plasmid DNA expression and pharmacokinetics.
Intratracheal administration of plasmid DNA resulted in gene expression in mouse airways in the absence of any enhancing agent. Administration of plasmid DNA encoding the chloramphenicol acetyltransferase gene (CAT) in sterile water lead to CAT transgene expression that peaked between 1 and 3 days and was detected up to 28 days after DNA administration. Transgene expression was independent of mouse gender, age and strain. Levels of expression from DNA in various isotonic solutions did not differ from levels obtained with DNA administered in water, suggesting that transfection is not dependent on damage to airway cells caused by a hypo-osmotic delivery vehicle. Pharmacokinetic studies using radiolabeled plasmid DNA showed that DNA was rapidly degraded, while higher levels of radioactivity were retained for longer duration following administration of cationic liposome-DNA complexes in the airway. Southern blot and PCR analysis confirmed that DNA complexed with DOTMA-DOPE was retained in the airways for a longer period. However, cationic liposomes DOTMA-DOPE (1:1) or DOTAP complexed with DNA, did not enhance expression over DNA alone. These results suggest that 'naked' plasmid DNA should be included as a control in all studies on intratracheal gene delivery using nonviral systems.
Meyer
K B
KB
School of Pharmacy, University of California, San Francisco 94143-0446, USA.
Thompson
M M
MM
Levy
M Y
MY
Barron
L G
LG
Szoka
F C
FC
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000493:pharmacokinetics / Q000494:pharmacology; D004337:Drug Carriers; D016503:Drug Delivery Systems; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D008081:Liposomes; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014132:Trachea / Q000201:enzymology*
D004337:Drug Carriers; D008081:Liposomes; D015500:Chloramphenicol O-Acetyltransferase