Mechanistic study of transfection of chitosan/DNA complexes coated by anionic poly(γ-glutamic acid)

Zi Xian Liao, Shu Fen Peng, Yi Cheng Ho, Fwu Long Mi, Barnali Maiti, Hsing Wen Sung

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Chitosan (CS) has been investigated as a non-viral carrier for gene delivery, but resulting in a relatively low transfection. To address this concern, we developed a ternary system comprised the core of CS/DNA complex and the outer coating of an anionic polymer, poly(γ-glutamic acid) (γ-PGA). In molecular dynamic (MD) simulations, we found that γ-PGA was entangle tightly with the excess CS emanating from the surface of test complexes, thus making them more compact. With γ-PGA coating, the extent of test complexes internalized and their transfection efficiency were evidently enhanced. Trypsin treatment induced a concentration-dependent decrease in internalization of the γ-PGA-coated complexes, suggesting a specific protein-mediated endocytosis. The endocytosis inhibition study indicates that the γ-glutamyl transpeptidase (GGT) present on cell membranes was responsible for the uptake of test complexes. The amine group in the N-terminal γ-glutamyl unit on γ-PGA played an essential role in the interaction with GGT. When entangled with CS, the free N-terminal γ-glutamyl unit of γ-PGA on test complexes was exposed and might thus be accommodated within the γ-glutamyl binding pocket of the membrane GGT. Above results suggest that the γ-PGA coating on CS/DNA complexes can significantly enhance their cellular uptake via a specific GGT-mediated pathway. Knowledge of the uptake mechanism is crucial for the development of an efficient vector for gene transfection.

Original languageEnglish
Pages (from-to)3306-3315
Number of pages10
JournalBiomaterials
Volume33
Issue number11
DOIs
Publication statusPublished - Apr 2012
Externally publishedYes

Fingerprint

Prostaglandins A
Chitosan
Transfection
Glutamic Acid
DNA
Acids
Coatings
Genes
Endocytosis
Ternary systems
Cell membranes
Molecular dynamics
Amines
gamma-Glutamyltransferase
Molecular Dynamics Simulation
Proteins
Membranes
Trypsin
Polymers
Computer simulation

Keywords

  • γ-glutamyl transpeptidase
  • γ-glutamyl unit
  • Gene transfection
  • Poly(γ-glutamic acid)
  • Receptor-mediated endocytosis

ASJC Scopus subject areas

  • Biomaterials
  • Bioengineering
  • Ceramics and Composites
  • Mechanics of Materials
  • Biophysics

Cite this

Mechanistic study of transfection of chitosan/DNA complexes coated by anionic poly(γ-glutamic acid). / Liao, Zi Xian; Peng, Shu Fen; Ho, Yi Cheng; Mi, Fwu Long; Maiti, Barnali; Sung, Hsing Wen.

In: Biomaterials, Vol. 33, No. 11, 04.2012, p. 3306-3315.

Research output: Contribution to journalArticle

Liao, Zi Xian ; Peng, Shu Fen ; Ho, Yi Cheng ; Mi, Fwu Long ; Maiti, Barnali ; Sung, Hsing Wen. / Mechanistic study of transfection of chitosan/DNA complexes coated by anionic poly(γ-glutamic acid). In: Biomaterials. 2012 ; Vol. 33, No. 11. pp. 3306-3315.
@article{c9d719da432f408ab06a6abaa9b2910b,
title = "Mechanistic study of transfection of chitosan/DNA complexes coated by anionic poly(γ-glutamic acid)",
abstract = "Chitosan (CS) has been investigated as a non-viral carrier for gene delivery, but resulting in a relatively low transfection. To address this concern, we developed a ternary system comprised the core of CS/DNA complex and the outer coating of an anionic polymer, poly(γ-glutamic acid) (γ-PGA). In molecular dynamic (MD) simulations, we found that γ-PGA was entangle tightly with the excess CS emanating from the surface of test complexes, thus making them more compact. With γ-PGA coating, the extent of test complexes internalized and their transfection efficiency were evidently enhanced. Trypsin treatment induced a concentration-dependent decrease in internalization of the γ-PGA-coated complexes, suggesting a specific protein-mediated endocytosis. The endocytosis inhibition study indicates that the γ-glutamyl transpeptidase (GGT) present on cell membranes was responsible for the uptake of test complexes. The amine group in the N-terminal γ-glutamyl unit on γ-PGA played an essential role in the interaction with GGT. When entangled with CS, the free N-terminal γ-glutamyl unit of γ-PGA on test complexes was exposed and might thus be accommodated within the γ-glutamyl binding pocket of the membrane GGT. Above results suggest that the γ-PGA coating on CS/DNA complexes can significantly enhance their cellular uptake via a specific GGT-mediated pathway. Knowledge of the uptake mechanism is crucial for the development of an efficient vector for gene transfection.",
keywords = "γ-glutamyl transpeptidase, γ-glutamyl unit, Gene transfection, Poly(γ-glutamic acid), Receptor-mediated endocytosis",
author = "Liao, {Zi Xian} and Peng, {Shu Fen} and Ho, {Yi Cheng} and Mi, {Fwu Long} and Barnali Maiti and Sung, {Hsing Wen}",
year = "2012",
month = "4",
doi = "10.1016/j.biomaterials.2012.01.013",
language = "English",
volume = "33",
pages = "3306--3315",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier Science Ltd",
number = "11",

}

TY - JOUR

T1 - Mechanistic study of transfection of chitosan/DNA complexes coated by anionic poly(γ-glutamic acid)

AU - Liao, Zi Xian

AU - Peng, Shu Fen

AU - Ho, Yi Cheng

AU - Mi, Fwu Long

AU - Maiti, Barnali

AU - Sung, Hsing Wen

PY - 2012/4

Y1 - 2012/4

N2 - Chitosan (CS) has been investigated as a non-viral carrier for gene delivery, but resulting in a relatively low transfection. To address this concern, we developed a ternary system comprised the core of CS/DNA complex and the outer coating of an anionic polymer, poly(γ-glutamic acid) (γ-PGA). In molecular dynamic (MD) simulations, we found that γ-PGA was entangle tightly with the excess CS emanating from the surface of test complexes, thus making them more compact. With γ-PGA coating, the extent of test complexes internalized and their transfection efficiency were evidently enhanced. Trypsin treatment induced a concentration-dependent decrease in internalization of the γ-PGA-coated complexes, suggesting a specific protein-mediated endocytosis. The endocytosis inhibition study indicates that the γ-glutamyl transpeptidase (GGT) present on cell membranes was responsible for the uptake of test complexes. The amine group in the N-terminal γ-glutamyl unit on γ-PGA played an essential role in the interaction with GGT. When entangled with CS, the free N-terminal γ-glutamyl unit of γ-PGA on test complexes was exposed and might thus be accommodated within the γ-glutamyl binding pocket of the membrane GGT. Above results suggest that the γ-PGA coating on CS/DNA complexes can significantly enhance their cellular uptake via a specific GGT-mediated pathway. Knowledge of the uptake mechanism is crucial for the development of an efficient vector for gene transfection.

AB - Chitosan (CS) has been investigated as a non-viral carrier for gene delivery, but resulting in a relatively low transfection. To address this concern, we developed a ternary system comprised the core of CS/DNA complex and the outer coating of an anionic polymer, poly(γ-glutamic acid) (γ-PGA). In molecular dynamic (MD) simulations, we found that γ-PGA was entangle tightly with the excess CS emanating from the surface of test complexes, thus making them more compact. With γ-PGA coating, the extent of test complexes internalized and their transfection efficiency were evidently enhanced. Trypsin treatment induced a concentration-dependent decrease in internalization of the γ-PGA-coated complexes, suggesting a specific protein-mediated endocytosis. The endocytosis inhibition study indicates that the γ-glutamyl transpeptidase (GGT) present on cell membranes was responsible for the uptake of test complexes. The amine group in the N-terminal γ-glutamyl unit on γ-PGA played an essential role in the interaction with GGT. When entangled with CS, the free N-terminal γ-glutamyl unit of γ-PGA on test complexes was exposed and might thus be accommodated within the γ-glutamyl binding pocket of the membrane GGT. Above results suggest that the γ-PGA coating on CS/DNA complexes can significantly enhance their cellular uptake via a specific GGT-mediated pathway. Knowledge of the uptake mechanism is crucial for the development of an efficient vector for gene transfection.

KW - γ-glutamyl transpeptidase

KW - γ-glutamyl unit

KW - Gene transfection

KW - Poly(γ-glutamic acid)

KW - Receptor-mediated endocytosis

UR - http://www.scopus.com/inward/record.url?scp=84856587267&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856587267&partnerID=8YFLogxK

U2 - 10.1016/j.biomaterials.2012.01.013

DO - 10.1016/j.biomaterials.2012.01.013

M3 - Article

VL - 33

SP - 3306

EP - 3315

JO - Biomaterials

JF - Biomaterials

SN - 0142-9612

IS - 11

ER -