Modified Tat peptide with cationic lipids enhances gene transfection efficiency via temperature-dependent and caveolae-mediated endocytosis

Seiichi Yamano, Jisen Dai, Carlo Yuvienco, Sachin Khapli, Amr Moursi, Jin Montclare

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)278-285
Number of pages8
JournalJournal of Controlled Release
Volume152
Issue number2
DOIs
StatePublished - Jun 10 2011

Fingerprint

Caveolae
Endocytosis
Transfection
Lipids
Peptides
Temperature
Genes
DNA
Filipin
Clathrin
Chlorpromazine
Chloroquine
Luciferases
Particle Size
Histidine
Cysteine
HIV-1
Plasmids
Cell Line
Serum

Keywords

  • Gene delivery
  • Lipids
  • Plasmid DNA
  • Tat peptide
  • Transfection

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Modified Tat peptide with cationic lipids enhances gene transfection efficiency via temperature-dependent and caveolae-mediated endocytosis. / Yamano, Seiichi; Dai, Jisen; Yuvienco, Carlo; Khapli, Sachin; Moursi, Amr; Montclare, Jin.

In: Journal of Controlled Release, Vol. 152, No. 2, 10.06.2011, p. 278-285.

Research output: Contribution to journalArticle

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