HIV-1 Tat protein transduction domain peptide facilitates gene transfer in combination with cationic liposomes

L. Hyndman, J. L. Lemoine, L. Huang, D. J. Porteous, A. C. Boyd, X. Nan

Research output: Contribution to journalArticlepeer-review

Abstract

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.
Original languageEnglish
Pages (from-to)435-444
Number of pages10
JournalJournal of Controlled Release
Volume99
Issue number3
Publication statusPublished - 2004

Keywords

  • 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

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