The protein transduction domain (PTD) of the HIV-1 Tat protein can facilitate the cellular and nuclear uptake of macromolecular particles. Here, we demonstrate that incorporation without covalent linkage of a 17-amino acid PTD peptide into gene delivery lipoplexes improves gene transfer. Tat/Liposome/DNA (TLD) transfection, as evaluated by Fluorescence Activated Cell Scan analysis of a Green Fluorescence Protein expression plasmid, enabled transfection of highly recalcitrant primary cells in the form of air/liquid interface cultures of sheep tracheal epithelium. Treatment with chloroquine increased, and incubation at low temperature decreased, TLD transfection, suggesting that the endocytosis uptake pathway is involved.
|Number of pages||10|
|Journal||Journal of Controlled Release|
|Publication status||Published - 2004|
- Amino Acid Sequence Animals Carrier Proteins/genetics/pharmacokinetics Cell Line, Tumor Chloroquine/pharmacology DNA/genetics/metabolism DNA-Binding Proteins/genetics/metabolism Deoxyribonuclease I/drug effects/physiology Drug Evaluation, Preclinical/methods Electrophoretic Mobility Shift Assay/methods Endocytosis/drug effects/physiology Fatty Acids, Monounsaturated/chemistry/pharmacokinetics Flow Cytometry/methods Forecasting Gene Products, tat/genetics/*pharmacokinetics Gene Therapy/methods Genetic Vectors/genetics/pharmacokinetics HIV-1/genetics/*physiology Humans Liposomes/chemistry/*pharmacokinetics Macromolecular Substances/chemistry Particle Size Phosphatidylethanolamines/pharmacokinetics Plasmids/chemistry/pharmacokinetics Quaternary Ammonium Compounds/chemistry/pharmacokinetics Sequence Analysis, Protein/methods Sheep Trachea/drug effects/pathology/physiology Transfection/*methods tat Gene Products, Human Immunodeficiency Virus