Gene transfer and analysis of the gene expression mechanism using pDNA/chitosan complexes coated with lactose-modified PEG derivatives

Rieko Okan, Kohei Koya, Yoshiyuki Koyama, Toshinori Sato

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Chitosan is one of naturally occurring polycations and is useful as a non-viral vector for gene delivery into cells. As PEG-C is carboxylic acid-pendant poly(ethylene glycol) derivatives, the negatively charged PEG-C can coat the surface of positively charged plasmid/chitosan complexes. The pDNA/chitosan complexes coated with the lactose modified PEG-C showed the high gene expression. To analyze the cell transfection mechanism, the amount of cell uptake was determined using a FACS, and intracellular transfer of the complexes was observed using a confocal laser scanning microscope (CLSM). The amount of cell uptake did not correlate with the gene expression, but the cell uptake of lactose-bearing complexes is faster than conventional PEG-C complexes. The CSLM showed that the pDNA/chitosan/PEG-C and pDNA/chitosan/Lac-PEG-C complexes were taken up into cells without dissociation. Moreover, the pDNA/chitosan and pDNA/chitosan/Lac-PEG-C complexes showed nuclear import after eight hours.

Original languageEnglish
Title of host publicationPolymer Preprints, Japan
Pages2145
Number of pages1
Volume54
Edition1
Publication statusPublished - 2005
Event54th SPSJ Annual Meeting 2005 - Yokohama, Japan
Duration: 2005 May 252005 May 27

Other

Other54th SPSJ Annual Meeting 2005
CountryJapan
CityYokohama
Period05/5/2505/5/27

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Keywords

  • Carbohydrate recognition
  • Chitosan
  • Confocal laser scanning microscope
  • FACS
  • Gene transfer
  • PEG

ASJC Scopus subject areas

  • Engineering(all)

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