Low Molecular Weight Branched PEI Binding to Linear DNA

• Utsuno Kuniharu

  Department of Science & Engineering for Materials, National Institute of Technology, Tomakomai College

• Kono Hiroyuki

  Department of Science & Engineering for Materials, National Institute of Technology, Tomakomai College

• Tanaka Emika

  Department of Science & Engineering for Materials, National Institute of Technology, Tomakomai College

• Jouna Nozomi

  Department of Science & Engineering for Materials, National Institute of Technology, Tomakomai College

• Kojima Yohichiro

  Department of Electrical & Electronic Engineering, Hokkaido University of Science

• Uludağ Hasan

  Department of Chemical & Materials Engineering, University of Alberta Department of Biomedical Engineering, University of Alberta Faculty of Pharmacy & Pharmaceutical Sciences, University of Alberta

抄録

Polyethylenimine (PEI) is one of the most versatile non-viral vectors used in gene therapy, especially for delivering plasmid DNA to human cells. However, a good understanding of PEI binding to DNA, the fundamental basis for the functioning of PEI as a vector, has been missing in the literature. In this study, PEI (branched, 600 Da) binding to DNA was examined by isothermal titration calorimetry (ITC), quartz crystal microbalance (QCM) and a complementary set of analysis tools. We demonstrated that a separation between the binding heat and the condensation heat is needed and that the excluded site model should be used for PEI binding stage in the ITC analysis. The equilibrium constant for PEI binding to DNA was determined to be 2.5×10⁵ M⁻¹ from the ITC analysis, and as 2.3×10⁵ M⁻¹ from the QCM analysis. Additionally, we suggested that the 600 Da branched PEI binds to the major groove of DNA and the rearrangement of PEI on DNA may be difficult to occur because of the small dissociation rate. The binding analysis presented here can be employed to improve our understanding of the functioning of PEI and PEI-like non-viral vectors.

収録刊行物

• ＣＨＥＭＩＣＡＬ　＆　ＰＨＡＲＭＡＣＥＵＴＩＣＡＬ　ＢＵＬＬＥＴＩＮ

  ＣＨＥＭＩＣＡＬ　＆　ＰＨＡＲＭＡＣＥＵＴＩＣＡＬ　ＢＵＬＬＥＴＩＮ 64 (10), 1484-1491, 2016

  公益社団法人 日本薬学会

キーワード

• DNA condensation
• polyethylenimine
• isothermal titration calorimetry
• quartz crystal microbalance
• thermodynamics
• kinetics

詳細情報

• CRID: 1390282679154412800
• NII論文ID: 130005598026
• NII書誌ID: AA00602100
• DOI: 10.1248/cpb.c16-00454
• ISSN: 13475223
• ISSN: 00092363
• NDL書誌ID: 027623429
• PubMed: 27725501
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I027623429
• Web Site: https://www.jstage.jst.go.jp/article/cpb/64/10/64_c16-00454/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
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