2006 Fiscal Year Final Research Report Summary
Molecular Design of DNA Conjugate and its Applications to Biosensing
| Project/Area Number |
15200039
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | RIKEN |
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Principal Investigator |
MAEDA Mizuo RIKEN, Bioengineering Laboratory, Chief Scientist, 前田バイオ工学研究室, 主任研究員 (10165657)
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| Co-Investigator(Kenkyū-buntansha) |
TAKARADA Tohru RIKEN, Bioengineering Laboratory, Research Scientist, 前田バイオ工学研究室, 研究員 (30336010)
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| Project Period (FY) |
2003 – 2006
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| Keywords | Nanoparticles / Colloids / Electrophoresis / DNA / Double helix / Gene diagnosis / Thermoresponsive polymers |
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| Research Abstract |
We developed novel biosensing technologies on the basis of our unique principle, and presented facile, effective and high-sensitive analytical methods. The principle consists in single-base discrimination by using non-cross-linking aggregation of DNA-carrying nanoparticles, which are formed through the self-assembly of thermoresponsive polymers grafted with oligodeoxyribonucleotides. In this study, we devised new DNA-conjugated materials, including DNA-responsive hydrogels, DNA-synthetic polymer block-type conjugates, and DNA-linked gold nanoparticles, as analytical tools for gene diagnostics based on this principle.
Achievements are summarized as follows:
1. The unique aggregation of DNA-immobilized polystyrene Latex and DNA-linked gold nanoparticles was demonstrated, indicating that the principle would hold in general.
2. Sequence-dependent, drastic change was found in electrophoretic mobility of DNA-carrying nanoparticles.
3. Novel hybrid hydrogels containing rationally designed single-stranded DNA as the cross-linker were developed to shrink or swell in response to sequence of sample DNA, allowing for precise discrimination of single base difference.
4. Affinity capillary electrophoresis using DNA-synthetic polymer block-type conjugates as affinity ligands was developed to separate single-stranded sample DNA and its point mutant.
5. Surface plasmon resonance imaging was conducted on a microchip for high-sensitive detection of DNA-linked gold nanoparticles deposited onto the surface in a non-cross-linking configuration.
6. Multiplex gene mutation analysis was realized on the basis of affinity electrophoresis in micro fluidic channels. With the aid of this research fund, we succeeded to construct biosensing systems by designing various DNA-conjugated materials.
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