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。 项目总结/摘要 适体是结合特定靶标（配体）的功能性寡核苷酸，所述靶标通常是小的寡核苷酸。 有机分子一些体外选择的RNA适体可以结合并实质上增加RNA的表达。 荧光配体的荧光量子产率，所述荧光配体在它们的未结合态中是弱荧光的 状态因此，开发可使用适体的方法具有相当大的兴趣 用于活细胞中分子的荧光成像。扩大这些方法的实用性和适应性 这些平台可能会导致与人类疾病相关的生物标志物的先进检测策略。 然而，限制在于每个RNA适体仅能够识别限定的， 有机分子的特征结构。在这里，我们描述（目标1）的设计师小的应用程序 分子，使一个单一的RNA适体，以检测多种以及结构多样的疾病- 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（目的2）通过提出以下建议来扩大RNA适体的翻译潜力： 一种将可重用性引入ABA系统的芯片设计。我们将微阵列固体表面与 纳米针和纳米管用于样本的捕获辅助检测。这些原理验证研究 将为开发新的分子工具奠定基础，以进一步研究细胞功能障碍， 导致即时护理应用。

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.