Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine

Enhanced cellular association by hydrophobic core formation

I. A. Khalil, S. Futaki, M. Niwa, Y. Baba, Noritada Kaji, H. Kamiya, H. Harashima

Research output: Contribution to journalArticle

138 Citations (Scopus)

Abstract

The internalization mechanisms associated with octaarginine and stearyl-octaarginine were investigated using confocal laser microscopy and flow cytometric analysis. Octaarginine is able to translocate through cell membranes in a manner that does not exactly involve the classical endocytic pathways of internalization. However, when a stearyl moiety is attached to the N-terminus of octaarginine, the internalization shifts mainly to an endocytosis-dependent pathway. The transfection efficiency of stearyl-octaarginine was significantly higher than that of octaarginin. To understand the mechanism of the improved gene transfer by the N-terminal stearylation of octaarginine, the gene transfer processes mediated by octaarginine or stearyl-octaarginine were compared. Both octaarginine and stearyl-octaarginine are able to carry plasmid DNA into cells. The amount of plasmid DNA internalized as well as that delivered to the nucleus was higher in the case of stearyl-octaarginine. Even though the internalization mechanisms of octaarginine and stearyl-octaarginine were different, their complexes with plasmid DNA were internalized via the same pathway, presumably, the clathrin-mediated pathway of endocytosis. The results of the atomic force microscopy revealed that stearyl-octaarginine, but not octaarginine, can completely condense the DNA into stable complexes that can be highly adsorbed to the cell surface and subsequently highly internalized. Therefore, using stearylated-octaarginine provided higher internalization of plasmid DNA into cells, due to enhanced cellular association, as well as higher nuclear delivery. The results presented in this study provide a better understanding of the mechanisms of improved transfection using stearylated-octaarginine. The concept of using stearylated peptides may aid in the development of more efficient nonviral gene vectors.

Original languageEnglish
Pages (from-to)636-644
Number of pages9
JournalGene Therapy
Volume11
Issue number7
DOIs
Publication statusPublished - Apr 1 2004
Externally publishedYes

Fingerprint

Genes
Plasmids
DNA
Endocytosis
Confocal Microscopy
Transfection
octaarginine
Clathrin
Atomic Force Microscopy
stearyl-octaarginine
Cell Membrane
Peptides

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine : Enhanced cellular association by hydrophobic core formation. / Khalil, I. A.; Futaki, S.; Niwa, M.; Baba, Y.; Kaji, Noritada; Kamiya, H.; Harashima, H.

In: Gene Therapy, Vol. 11, No. 7, 01.04.2004, p. 636-644.

Research output: Contribution to journalArticle

Khalil, I. A. ; Futaki, S. ; Niwa, M. ; Baba, Y. ; Kaji, Noritada ; Kamiya, H. ; Harashima, H. / Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine : Enhanced cellular association by hydrophobic core formation. In: Gene Therapy. 2004 ; Vol. 11, No. 7. pp. 636-644.
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