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

Design of tailor-made biosensors for cellular and chip applications

为细胞和芯片应用定制生物传感器的设计

基本信息

批准号：
15310147
负责人：
MORII Takashi
金额：
$ 8.9万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15310147/
关键词：
BIOSENSOR RECEPTOR SECOND MESSENGERS BOMBINATORIAL CHEMISTRY MOLECULAR RECOGNITION STRUCTURE-BASED DESIGN PEPTIDE RNA

项目摘要

The tailor-made receptors and enzymes are comprised of two subunits designed by the structure-based method and their functions are optimized through the combinatorial approach. We have designed a new class of scaffold for tailor-made receptors, in which a short peptide and RNA with randomized nucleotide region form a stable and specific complex In vitro selection of RNA oligonucleotides from the randomized "ribonucleopeptide (RNP)" pool afforded RNP receptors specific for ATP.In this research, we have expanded the utility of ribonucleopeptides to tailoring fluorescent biosensors. Construction of fluorescent biosensors with desired characteristics, i.e, high signal-to-noise ratios, detection wavelengths and concentration ranges for ligand detection, is not a straight forward task. Fluorescent RNP sensors for nucleotide triphosphates are constructed by two library selection steps. The first step tailors RNP receptors specific for a ligand by in vitro selection from an RNA-diverged RNP li … More brary. In the second step, combination of the RNA subunits from the first step and a series of fluorophore-modified peptide subunits affords fluorescent RNP libraries, from which RNP sensors with desired optical sensing properties are selected in a high-throughput manner. Screening of the fluorescence emission intensities in the presence of increasing concentrations of ATP allowed titration analysis of the fluorescent RNP library, which enabled a selection of ATP sensors responding within certain concentration ranges of ATP.The fluorescent sensor would be an ideal tool for the convenient measurements of cellular second messengers. We have designed a protein based sensor for IP_3 by exploring the selective IP_3 binding properties of pleckstrin homology (PH) domain. Several fluorescent sensors for inositol-1,3,4,5-tetrakisphosphate were obtained in this research. The strategy developed here afford functional small proteins essential for the nano-biotechnology tools such as biosensors and protein chips. Less

通过基于结构的方法设计了两个亚基，并通过组合方法优化了它们的功能。我们设计了一种新的受体支架，其中一个短肽和具有随机核苷酸区域的RNA形成一个稳定的和特异性的复合物。从随机的“核糖核肽（RNP）”库中体外选择RNA寡核苷酸，得到对ATP特异的RNP受体。构建具有所需特性的荧光生物传感器，即高信噪比、检测波长和用于配体检测的浓度范围，不是一个简单的任务。通过两个文库选择步骤构建用于核苷酸三磷酸的荧光RNP传感器。第一步是通过体外选择从RNA趋异的RNP中定制对配体特异的RNP受体。 ...更多信息 - 是的在第二步中，来自第一步的RNA亚基和一系列荧光团修饰的肽亚基的组合提供了荧光RNP文库，从该文库中以高通量方式选择具有所需光学传感特性的RNP传感器。在增加浓度的ATP存在下的荧光发射强度的筛选允许对荧光RNP文库进行滴定分析，这使得能够选择在一定浓度范围内响应的ATP传感器。我们利用pleckstrin同源结构域（PH）对IP_3的选择性结合特性，设计了一种基于蛋白质的IP_3传感器。本研究获得了几种肌醇-1，3，4，5-四磷酸荧光传感器。该策略为生物传感器和蛋白质芯片等纳米生物技术工具提供了必需的功能性小蛋白。少