Sensing in Living Cells: Expressable RNA-based FRET Probes

在过去的几年里，我们对小核糖核酸(RNAs)在控制细胞内过程中所起的重要作用的了解有了极大的增长。这个结合了化学和分子生物学的跨学科项目的目标是为细胞内的小RNA增加额外的人工功能：荧光报告细胞内的过程。为此，我们将设计新的小RNA，既能对代谢物做出反应，又能结合发光染料(与受体结合后增加荧光的染料)。新的代谢物荧光探针将有助于增加我们对细胞内过程的了解，从而立即带来基础科学上的好处，并对人类健康产生更长期的影响。博士后科学家或研究生、本科生和高中生都将接触到多学科方法，并将学习如何开发和优化有用的细胞内试剂。我们将用来演示新的基于RNA的探针的功能的生物之一是社会阿米巴虫，即网柄蠕虫；因此，我们可能会制作出极具视觉吸引力的电影，用来激发参与其中的K-12学生对科学的兴趣，并解释多细胞、进化、寄生、社会行为、利他主义和作弊的起源等基本原理。

Over the past several years our understanding of the important role that small ribonucleic acids (RNAs) play in controlling intracellular processes has grown tremendously. The goal of this interdisciplinary project, combining chemistry and molecular biology, is to add an additional artificial function to the intracellular small RNAs: the fluorescent reporting of intracellular process. For this purpose, we will engineer new small RNAs that will be both responsive to metabolites and bind light-up dyes (dyes that that increase fluorescence upon binding to the receptor). New fluorescent probes for metabolites would help increase our understanding of intracellular processes, leading to immediate basic science benefits and longer-term impact on human health. A postdoctoral scientist or a graduate student, undergraduates, and high?]school students will all be exposed to the multidisciplinary approach and will learn how to develop and optimize useful intracellular reagents. One of the organisms we will use to demonstrate the function of new RNA-based probes is the social amoebae, Dictyostellium; thus, we may produce highly visually attractive movies that will be used to spark the interest of participating K-12 students in science and to explain basic principles of multicellularity, evolution, parasitism, social behavior, origins of altruism and cheating.