title: Modified Tat peptide with cationic lipids enhances gene transfection efficiency via temperature-dependent and caveolae-mediated endocytosis.
abstract: The HIV-1 Tat peptide has been successfully used for intracellular gene delivery. Likewise, various lipid-based methods have shown increased endocytosis and can influence endosomal escape. This study combines the favorable properties of Tat peptide with that of lipid systems for DNA delivery. We combined the lipid FuGENE HD (FH) with the Tat peptide sequence modified with histidine and cysteine residues (mTat). mTat/FH transfection was evaluated by luciferase expression plasmid in five cell types. mTat/FH produced significant improvement in transfection efficiency of all cell lines when compared to FH or mTat. Treatment with chloroquine, associated with energy-dependent endocytosis, significantly increased transfection efficiency with mTat/FH while incubation at low temperature decreased it. The zeta potential of mTat/FH/DNA was significantly higher compared to FH, mTat, or their DNA combination in the presence of serum, and it was correlated with transfection efficiency. The particle size of the FH/DNA complex was significantly reduced by addition of mTat. Filipin III, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/FH transfection, but transfection was increased by chlorpromazine, an inhibitor of clathrin-mediated endocytosis. These findings demonstrated the feasibility of using a combination of mTat with lipids, utilizing temperature-dependent and caveolae-mediated endocytosis, as a potentially attractive non-viral gene vector.
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, 345 East 24th Street, New York, NY 10010, USA. sy23@nyu.edu
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yamano
authors.forename: Jisen
authors.initials: J
authors.lastname: Dai
authors.forename: Carlo
authors.initials: C
authors.lastname: Yuvienco
authors.forename: Sachin
authors.initials: S
authors.lastname: Khapli
authors.forename: Amr M
authors.initials: AM
authors.lastname: Moursi
authors.forename: Jin K
authors.initials: JK
authors.lastname: Montclare
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002412:Cations / Q000737:chemistry / Q000378:metabolism; D021941:Caveolae / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D002966:Clathrin / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014162:Transfection / Q000379:methods; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002412:Cations; D002966:Clathrin; D008055:Lipids; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; D004247:DNA