Antisense oligonucleotides bound in the polysaccharide complex and the enhanced antisense effect due to the low hydrolysis

Masami Mizu, Kazuya Koumoto, Takahisa Anada, Kazuo Sakurai, Seiji Shinkai

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Schizophyllan is a β-(1→3)-D-glucan and can form a novel complex with some single-chains of DNAs. As the preceding paper revealed, the polynucleotide bound in the complex is more stable to nuclease-mediated hydrolysis than the polynucleotide itself (i.e., naked polynucleotide). This paper examined possibility to apply this complex to an antisense DNA carrier, using an in vitro (cell-free) transcription/translation assay. In this assay, we used a plasmid DNA coding a green fluorescence protein (GFP) and an antisense DNA designed to hybridize the ribosome-binding site in the GFP-coded mRNA. When the antisense DNA was administered as the complex, a lower GFP expression efficiency (or higher antisense effect) is observed over naked DNA. This is because the antisense DNA in the complex is protected from the attack of deoxyribonuclease. When exonuclease I, which specifically hydrolyzes single DNA chains, was present in the GEP assay system, the antisense effect was not changed for the complex while being weakened in the naked antisense DNA system. These results imply that the exonuclease I cannot hydrolyze the antisense DNA in the complex, while it can hydrolyze naked DNA to reduce its antisense effect.

Original languageEnglish
Pages (from-to)3117-3123
Number of pages7
JournalBiomaterials
Volume25
Issue number15
DOIs
Publication statusPublished - 2004 Jul

Keywords

  • Antisense
  • Complexation
  • DNA
  • Gene expression
  • Polysaccharide

ASJC Scopus subject areas

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

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