Fluorescent nucleosides, nucleotides and oligonucleotides

项目总结 核酸及其构件在所有细胞活动中发挥着核心作用，因此具有巨大的 对疾病出现的影响，进而对人类健康的影响。研究这样的事件是复杂的 天然核酸库的非发射性质，这经常使研究人员无法使用 现代基于荧光的技术。忠实的最小扰动发出的核苷替代品可以因此 促进监测核苷、核苷酸和核酸在核苷/潮汐中的转化- “解决”，并推进基础研究、诊断工具和药物发现工作。 该计划的目标是设计和合成新的同象发射核苷和 核苷酸类似物，并将其用作监测核苷和基于核苷酸的探针 转化以及核酸的功能、结构、动力学和识别。具体地说，少校 将应对当代挑战，试图弥合重大差距，其中包括：(A)强大 生物物理技术，如荧光检测圆二色谱(FDCD)，引入了近五个 (B)多光子、成像和单分子光谱学- 基于实验，使用天然或最小扰动的寡核苷酸或核苷酸辅因子，严重 未得到充分利用；(C)同样，核苷/潮汐处理酶的单分子酶学没有 高级；(D)用于实时探索基本过程的探头，如肽基转移酶、相 缺乏分离的无膜细胞器的形成和信使核糖核酸的衰变；(E)核苷/潮基 代谢过程和基于核苷酸的信号事件不能被直接监测；和(F)高 核苷和核苷加工酶的吞吐量筛选不能实时进行 并且以高通量的方式进行，而无需使用忠实的发光替代衬底。 利用我们实验室开发的几个有用的发射核苷替代品家族，我们 将进一步提炼我们的“设计者”发射和同构的核苷/潮汐，并将它们应用于 解决上述挑战的办法。我们将追求新的物理和技术的进步 生物化学方法以及有效的实时筛查和诊断工具。这些努力将会扩大 社区的发射功能探测器武器库，推动未来在发现和成像方面的进展 申请。这些创新反过来又将进一步从根本上理解关键的生物过程 与疾病发展有关，并将对改善人类健康产生长期影响。

PROJECT SUMMARY Nucleic acids and their building blocks play central roles in all cellular events and, as such, have immense impact on the emergence of diseases and, in turn, on human health. Studying such events is complicated by the non-emissive nature of the natural nucleobases, which frequently deprives researchers from the use of modern fluorescence-based techniques. Faithful minimally perturbing emissive nucleoside surrogates can thus facilitate the monitoring of nucleoside, nucleotides and nucleic acids-based transformations at nucleoside/tide- “resolution”, and advance basic research, diagnostic tools and drug discovery efforts. The goal of the proposed program is to design and synthesize new isomorphic emissive nucleoside and nucleotide analogs and implement them as probes for monitoring nucleoside- and nucleotide-based transformations as well as nucleic acids function, structure, dynamics and recognition. Specifically, major contemporary challenges will be tackled in an attempt to bridge major gaps, among them: (a) Powerful biophysical techniques, such as Fluorescence-Detected Circular Dichroism (FDCD), introduced nearly five decades ago, remains practically unexplored; (b) Multiphoton, imaging and single molecule spectroscopy- based experiments, using native or minimally perturbed oligonucleotides or nucleotide cofactors, are severely underutilized; (c) Similarly, single molecule enzymology of nucleoside/tide processing enzymes has not advanced; (d) Probes for real time exploration of fundamental processes such as peptidyl transferase, phase separated membrane-less organelle formation and mRNA decay are lacking; (e) Nucleoside/tide-based metabolic processes and nucleotide-based signaling events cannot be directly monitored; and (f) High throughput screening for nucleosides and nucleosides processing enzymes cannot be performed in real-time and in a high throughput manner without the use of faithful emissive surrogate substrates. Capitalizing on several useful families of emissive nucleoside surrogates developed in our laboratory, we will further refine our “designer” emissive and isomorphic nucleosides/tides and apply them to advance solutions to the challenges articulated above. We will pursue the advancement of new physical and biochemical methods, as well as effective real-time screening and diagnostic tools. These efforts will expand the community's arsenal of emissive functional probes, driving future strides into discovery and imaging applications. These innovations, in turn, will further fundamental understanding of key biological processes related to disease development and will have long-term impact on improving human health.