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.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Carrier Proteins / genetics
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Carrier Proteins / pharmacokinetics
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Cell Line, Tumor
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Chloroquine / pharmacology
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DNA / genetics
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DNA / metabolism
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Deoxyribonuclease I / drug effects
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Deoxyribonuclease I / physiology
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Drug Evaluation, Preclinical / methods
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Electrophoretic Mobility Shift Assay / methods
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Endocytosis / drug effects
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Endocytosis / physiology
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Fatty Acids, Monounsaturated / chemistry
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Fatty Acids, Monounsaturated / pharmacokinetics
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Flow Cytometry / methods
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Forecasting
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Gene Products, tat / genetics
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Gene Products, tat / pharmacokinetics*
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Genetic Therapy / methods
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Genetic Vectors / genetics
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Genetic Vectors / pharmacokinetics
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HIV-1 / genetics
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HIV-1 / physiology*
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Humans
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Liposomes / chemistry
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Liposomes / pharmacokinetics*
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Macromolecular Substances / chemistry
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Particle Size
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Phosphatidylethanolamines / pharmacokinetics
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Plasmids / chemistry
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Plasmids / pharmacokinetics
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Quaternary Ammonium Compounds / chemistry
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Quaternary Ammonium Compounds / pharmacokinetics
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Sequence Analysis, Protein / methods
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Sheep
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Trachea / drug effects
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Trachea / pathology
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Trachea / physiology
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Transfection / methods*
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tat Gene Products, Human Immunodeficiency Virus
Substances
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Carrier Proteins
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DNA-Binding Proteins
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Fatty Acids, Monounsaturated
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Gene Products, tat
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Liposomes
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Macromolecular Substances
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Phosphatidylethanolamines
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Quaternary Ammonium Compounds
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tat Gene Products, Human Immunodeficiency Virus
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1,2-dielaidoylphosphatidylethanolamine
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Chloroquine
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DNA
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Deoxyribonuclease I
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1,2-dioleoyloxy-3-(trimethylammonium)propane