Activity-Based Aptamers as Bioimaging Tools for Probing Organics, Inorganics, and Enzymes

基于活性的适体作为探测有机物的生物成像工具， 无机物和酶。 项目摘要/摘要 适配子是与特定的靶(配体)结合的功能性寡核苷酸，通常是一个小的 有机分子。一些体外选择的RNA适配子可以结合并显著增加 未结合时荧光较差的荧光配体的荧光量子产率 州政府。因此，人们对开发可以使用适配子的方法非常感兴趣 用于对活细胞中的分子进行荧光成像。将这些方法扩展为实用和适应性强的方法 平台可能导致对与人类疾病相关的生物标记物的先进检测战略。 然而，局限性在于每个RNA适配子只能识别定义的， 有机分子的特征结构。这里我们描述(目标1)Designer Small的一个应用程序 使单个RNA适配子能够检测多种以及结构多样化的疾病的分子- 相关的生物标志物，包括无机分子和酶；我们称之为基于活性的适体 (ABA)系统。此外，我们将(目标2)通过提出以下建议来扩大RNA适配子的翻译潜力 将可重用性引入ABA系统的芯片设计。我们将用微阵列技术将固体表面 用于捕获辅助检测样品的纳米针和纳米吸管。这些原则证明研究 将为开发新的分子工具进一步研究细胞功能障碍和 导致使用护理点应用程序。

Activity-Based Aptamers as Bioimaging Tools for Probing Organics, Inorganics, and Enzymes. PROJECT SUMMARY / ABSTRACT Aptamers are functional oligonucleotides that bind a specific target (ligand), which is typically a small organic molecule. Some in vitro-selected RNA aptamers can bind and substantially increase the fluorescence quantum yield of fluorogenic ligands that are otherwise poorly fluorescent in their unbound state. Consequently, there is considerable interest in developing methods in which aptamers can be used for fluorescence imaging of molecules in living cells. Expansion of these methods as practical and adaptable platforms could lead to advanced detection strategies for biomarkers associated with human diseases. However, the limitations lie in that each RNA aptamer is only capable of recognizing a defined, characteristic structure of organic molecules. Here we describe (Aim 1) an application of designer small molecules to enable a single RNA aptamer to detect multiple as well as structurally diverse disease- associated biomarkers, including inorganic molecules and enzymes; we term this activity-based aptamer (ABA) system. In addition, we will (Aim 2) expand the translational potential of RNA aptamers by proposing a chip design that introduces reusability into ABA systems. We will microarray solid surfaces with nanoneedles and nanostraws for capture-assisted detection of specimens. These proof-of-principle studies will lay the foundation for developing novel molecular tools to further investigate cellular dysfunction and lead to point-of-care application.