title: Long-term efficient gene delivery using polyethylenimine with modified Tat peptide.
abstract: Polyethylenimine (PEI), a cationic polymer, has been widely studied and shown great promise as an efficient gene delivery vehicle. Likewise, the HIV-1 Tat peptide, a cell-permeable peptide, has been successfully used for intracellular gene delivery. To improve the favorable properties of these two vectors, we combine PEI with the modified Tat peptide sequence bearing histidine and cysteine residues (mTat). In vitro mTat/PEI-mediated transfection was evaluated by luciferase expression plasmid in two cell types. mTat/PEI produced significant improvement (≈5-fold) in transfection efficiency of both cell lines with little cytotoxicity when compared to mTat alone, PEI alone, or four commercial reagents. The particle size of mTat/PEI/DNA complex was significantly smaller than mTat or PEI alone, and it was correlated with higher transfection efficiency. Filipin III, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/PEI transfection. In contrast, chlorpromazine, an inhibitor of clathrin-mediated endocytosis, did not. This suggested caveolae-mediated endocytosis as the transfection mechanism. Furthermore, the results of in vivo studies showed that animals administered mTat/PEI/DNA intramuscularly had significantly higher and longer luciferase expression (≈7 months) than those with mTat/DNA, PEI/DNA, or DNA alone, without any associated toxicity. The combination of mTat with PEI could significantly improve transfection efficiency, expanding the potential use as a non-viral gene vector both in vitro and in vivo.
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States. Electronic address: sy23@nyu.edu.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yamano
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Jisen
authors.initials: J
authors.lastname: Dai
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Hanatani
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Ken
authors.initials: K
authors.lastname: Haku
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Takuto
authors.initials: T
authors.lastname: Yamanaka
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Mika
authors.initials: M
authors.lastname: Ishioka
authors.affiliation: Department of Prosthodontics, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Tadahiro
authors.initials: T
authors.lastname: Takayama
authors.affiliation: Department of Chemical and Biomolecular Engineering, Polytechnic Institute of New York University, Brooklyn, NY 11201, United States.
authors.forename: Carlo
authors.initials: C
authors.lastname: Yuvienco
authors.affiliation: Division of Engineering, New York University Abu Dhabi, Abu Dhabi, United Arab Emirates.
authors.forename: Sachin
authors.initials: S
authors.lastname: Khapli
authors.affiliation: Department of Pediatric Dentistry, New York University College of Dentistry, New York, NY 10010, United States.
authors.forename: Amr M
authors.initials: AM
authors.lastname: Moursi
authors.affiliation: Department of Chemical and Biomolecular Engineering, Polytechnic Institute of New York University, Brooklyn, NY 11201, United States.
authors.forename: Jin K
authors.initials: JK
authors.lastname: Montclare
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D015696:Gene Products, tat / Q000008:administration & dosage* / Q000737:chemistry; D007273:Injections, Intramuscular; D051379:Mice; D010316:Particle Size; D010455:Peptides / Q000008:administration & dosage* / Q000737:chemistry; D011094:Polyethyleneimine / Q000008:administration & dosage*; D014162:Transfection / Q000379:methods*; D019076:Transgenes
chemical_list: D015696:Gene Products, tat; D010455:Peptides; D011094:Polyethyleneimine
keywords: Gene delivery; Non-viral vector; Plasmid DNA; Polyethylenimine; Tat peptide; Transfection