| Project/Area Number |
13490021
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
広領域
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| Research Institution | Kyoto University |
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Principal Investigator |
AOYAMA Yasuhiro Kyoto University, Graduate School of Engineering, Professor, 工学研究科, 教授 (00038093)
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| Co-Investigator(Kenkyū-buntansha) |
SANDO Shinsuke Kyoto University, Graduate School of Engineering, Research Associate, 工学研究科, 助手 (20346084)
AKIYOSHI Kazunari Tokyo Medical and Dental University, Institute of Biomaterials and Bioengineering, Professor, 生体材料工学研究所, 教授 (90201285)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥16,000,000 (Direct Cost: ¥16,000,000)
Fiscal Year 2003: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥8,800,000 (Direct Cost: ¥8,800,000)
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| Keywords | gene / gene delivery / artificial glycovirus / glycocluster / galactose / cell recognition / proteoglycan / cell adhesion / 人工ウイルス / 肝細胞 / エンドサイトーシス / 量子ドット / コンドロイチン硫酸 / 遺伝子診断 / ナノパーティクル / 水素結合 / 核酸 / 遺伝子導入 / 細胞 / トランスフェクション / 薬物運搬 / ルシフェラーゼ / 受容体 |
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| Research Abstract |
We constructed a gene delivery system using a calix[4]resorcarene-based glycocluster compounds as hosts/carriers. The major achievements are as follows.
(1) Glycoclusters form micelle-like nanoparticles, which are agglutinated with phosphate ions. This is entropy-driven via dehydration of glycoclusters and anions.
(2) Glycolcusters also strongly bind to plasmid DNAs to give "glycoviruses" having a size of ca. 50 nm. They are capable of transfection of various cells via endocytosis to express encoded proteins.
(3) Endocytosis is size-dependent and is optimized at around 50 nm. Glycoviruses having terminal galactose residues undergo further aggregation to lower the transfection activities. On the other hand, they are capable of targeting the hepatic cells via receptor-mediated specific pathway. The overall activity, however, is kept low due to the size factor.
(4) The partially galactose-functionalized glyovirus is free from aggregation and show a high affinity as well selectivity to the hepatic cells both in vitro and in vivo (mouse).
(5) We are thus successful in constructing a highly hepatocyte-targeting gene delivery system using a non-aggregating galactose-virus having a manipulated mode of galactose functionalization.
(6) In view of the remarkable roles of proteoglycans in the extracellular matrices, we also prepared condroitin-sulfate functionalized macrocyclic clusters. They show a remarkably potent inhibition effect toward fibronectin-integrin mediated cell adhesion, probably as a result of stabilization of protein complexes from undergoing aggregation
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