Linear double-stranded DNA that mimics an infective tail of virus genome to enhance transfection

Takahisa Anada, Ryouji Karinaga, Kazuya Koumoto, Masami Mizu, Takeshi Nagasaki, Yoshio Kato, Kazunari Taira, Seiji Shinkai, Kazuo Sakurai

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Our previous work showed that a natural β-(1 → 3)-d-glucan schizophyllan (SPG) can form a stable complex with single-stranded oligonucleotides (ssODNs). When protein transduction peptides were attached to SPG and this modified SPG was complexed with ssODNs, the resultant complex could induce cellular transfection of the bound ODNs, without producing serious cytotoxicity. However, no technique was available to transfect double-stranded DNAs (dsDNA) or plasmid DNA using SPG. This paper presents a new approach to transfect dsDNA, showing preparation and transfection efficiency for a minimal-size gene having a loop-shaped poly(dA)80 on both ends. This poly(dA) loops of dsDNA can form a complex with SPG. An siRNA-coding dsDNA with the poly(dA) loop was complexed with Tat-attached SPG to silence luciferase expression. When LTR-Luc-HeLa cells that can express luciferase under the control of the LTR promoter were exposed to this complex, the expression of luciferase was suppressed (i.e., RNAi effect was enhanced). Cytotoxicity studies showed that the Tat-SPG complex induced much less cell death compared to polyethylenimine, indicating that the proposed method caused less harm than the conventional method. The Tat-SPG/poly(dA) looped dsDNA complex had a structure similar to the viral genome in that the dsDNA ends were able to induce transfection and protection. The present work identifies the SPG and poly(dA) looped minimum-sized gene combination as a candidate for a non-toxic gene delivery system.

Original languageEnglish
Pages (from-to)529-539
Number of pages11
JournalJournal of Controlled Release
Volume108
Issue number2-3
DOIs
Publication statusPublished - 2005 Nov 28

Keywords

  • Gene delivery
  • Minimum-sized gene transfer vector
  • Polysaccharide/DNA complex
  • Schizophyllan

ASJC Scopus subject areas

  • Pharmaceutical Science

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