2002 Fiscal Year Final Research Report Summary
Polycationic copolymers having stabilizing activity toward triplex DNA
| Project/Area Number |
12480260
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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 |
Biomedical engineering/Biological material science
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
MARUYAMA Atsushi Graduate School of Bioscience and biotechnology, Associate Professor, 大学院・生命理工学研究科, 助教授 (40190566)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEI Yoshiyukii Juntendo University, School of Medicine, Lecturer, 医学部, 講師 (10306954)
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| Project Period (FY) |
2000 – 2002
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| Keywords | Graft copolymers / Polycation / Oligonucleotides / DNA triplex / Gene expression / Polyion complex / Transcription factor / Collagen |
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| Research Abstract |
Stabilization of nucleotide hybridization is considered important for improving gene therapy using oligonucleotides. We have designed comb-type copolymer consisting of polycation backbone (polylysine) and hydrophilic side chains as a stabilizer for double and triple helical DNAs. The copolymer considerably increased the thermal stability of triple helical structure but did not affect the reversible transition between triple helical and single-stranded DNA. An in vitro electrophoretic mobility shift assay revealed that the copolymer remarkably increased association constants of both Hoogsteen and reverse Hoogsteen-type triple helix formation. Moreover the triple helix-stabilizing efficiency of the copolymer was significantly higher than that of other oligocations like spermine and spermidine. Not only being good DNA triple helix stabilizer, it has also been shown to accelerate DNA strand exchange reactions between double helical DNA and its complementary oligonucleotides. From these, we conclude that this copolymer is capable of either stabilizing or activating DNA hybrids, and may useful for gene targeting employing oligonucleotides.
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