Fluorescent Nucleosides and Oligonucleotides with New Capabilities

在这个由化学部门化学结构、动态和机制B项目资助的项目中，圣地亚哥州立大学的Byron Purse教授正在寻求对核碱基类似物（DNA和RNA的合成构建块）的化学结构、局部分子环境和荧光特性之间的关系进行深入的理解。这些荧光核碱基类似物（FBAs）是一类重要的分子探针，用于了解控制基因表达和遗传密码的维持和复制的生化机制。为这些应用设计荧光分子探针是具有挑战性的，而且FBAs的能力有限，因为还不可能预测当探针放置在生物环境中时它们的荧光会如何变化。本研究中不可或缺的是培养研究生的计划，他们将领导实验室工作并参与项目规划、数据分析、解释、合作和结果传播的各个方面。包括本科生研究人员的参与，以及SDSU物理化学实验室的基于课程的本科生研究经验（CURE），以进一步扩大参与。本项目有三个具体的技术目标：(1)开发具有高亮度和光稳定性的荧光核碱基类似物（FBAs），并在单分子水平上评估其在核酸荧光研究中的性能；(2)利用光谱特征分析和FRET发展FBA对同时测量短期和长期结构和动力学的能力；(3)利用新的模拟设计和合成、荧光研究和计算相结合，发展一种理论方法来理解FBA光物理和对局部环境的响应。该项目支持的研究旨在测量和了解当FBAs被放置在DNA和RNA的不同环境中时，荧光是如何变化的，并利用这种理解，与合成有机化学一起，创造出具有增强能力的新型荧光探针，以加速发现。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

In this project, funded by the Chemical Structure, Dynamic & Mechanism B Program of the Chemistry Division, Professor Byron Purse of San Diego State University is seeking to develop a deep understanding of the relationship between chemical structure, local molecular environment, and fluorescent properties of nucleobase analogues, synthetic building blocks for DNA and RNA. These fluorescent nucleobase analogues (FBAs) are an important class of molecular probes, used to understand the biochemical mechanisms that control gene expression and the maintenance and copying of the genetic code. Designing fluorescent molecular probes for these applications is challenging and the capabilities of FBAs are limited because it is not yet possible to predict how their fluorescence will change when the probes are placed in a biological context. Integral to this research is a plan to train graduate students who will lead the laboratory work and participate in all aspects project planning, data analysis, interpretation, collaborations, and the dissemination of results. The involvement of undergraduate researchers is included, as is a Course-based Undergraduate Research Experience (CURE) in the SDSU Physical Chemistry Lab to further broaden participation.This project has three specific technical objectives: (1) to develop fluorescent nucleobase analogues (FBAs) with high brightness and photostability and assess their performance in fluorescence studies of nucleic acids at the single-molecule level; (2) to develop the ability of pairs of FBAs to measure short and long-range structure and dynamics simultaneously using spectral feature analysis and FRET and (3) to develop a theoretical approach to understanding FBA photophysics and responses to local environment using a combination of new analogue design and synthesis, fluorescence studies, and computation. The research supported by this project seeks to measure and understand how fluorescence changes when FBAs are placed in differing contexts in DNA and RNA and use this understanding, together with synthetic organic chemistry, to create new fluorescent probes with enhanced capabilities to accelerate discovery.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.