权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular Design of DNA Conjugate and its Applications to Biosensing

DNA 缀合物的分子设计及其在生物传感中的应用

基本信息

批准号：
15200039
负责人：
MAEDA Mizuo
金额：
$ 31.45万
依托单位：
RIKEN
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15200039/
关键词：
Nanoparticles Colloids Electrophoresis DNA Double helix Gene diagnosis Thermoresponsive polymers

项目摘要

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.

我们基于独特的原理开发了新型生物传感技术，并提出了简便、有效、高灵敏度的分析方法。其原理在于通过使用携带 DNA 的纳米颗粒的非交联聚集来进行单碱基区分，这些纳米颗粒是通过接枝寡脱氧核糖核苷酸的热响应聚合物的自组装形成的。本研究基于这一原理，设计了新型DNA结合材料，包括DNA响应水凝胶、DNA合成聚合物嵌段型结合物和DNA连接金纳米粒子，作为基因诊断的分析工具。主要研究成果如下： 1．证明了DNA固定聚苯乙烯乳胶和DNA连接的金纳米粒子的独特聚集，表明该原理是普遍成立的。 2．携带DNA的纳米颗粒的电泳迁移率发生了序列依赖性的剧烈变化。3．新型混合水凝胶含有合理设计的单链DNA作为交联剂，可根据样本DNA序列收缩或膨胀，从而精确区分单碱基差异。4．建立了以DNA合成聚合物嵌段型缀合物为亲和配体的亲和毛细管电泳技术，用于分离单链样品DNA及其点突变体。 5．在微芯片上进行表面等离子体共振成像，以高灵敏度检测以非交联配置沉积在表面上的 DNA 连接金纳米粒子。 6．基于微流体通道亲和电泳实现了多重基因突变分析。借助这笔研究基金，我们通过设计各种DNA共轭材料，成功构建了生物传感系统。