Fluorescent nucleosides, nucleotides and oligonucleotides

荧光核苷、核苷酸和寡核苷酸

• 批准号: 10083552
• 负责人: YITZHAK TOR
• 金额: $ 46.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10083552
• 关键词: Automobile Driving; Basic Science; Biochemical; Biological Process; Circular Dichroism; Communities; DNA; Development; Diagnostic; Disease; Enzymatic Biochemistry; Enzymes; Event; Family; Fluorescence; Future; Goals; Health; Human; Image; Laboratories; Membrane; Metabolism; Methods; Modernization; Monitor; Nature; Nucleic Acids; Nucleosides; Nucleotides; Oligonucleotides; Organelles; Peptidyltransferase; Phase; Play; Process; RNA; Research Personnel; Resolution; Role; Signal Transduction; Spectrum Analysis; Structure; Techniques; Time; base; biophysical techniques; cofactor; design; drug discovery; experimental study; high throughput screening; improved; innovation; mRNA Decay; multiphoton imaging; nucleobase; nucleoside analog; nucleotide analog; programs; single molecule; tool; tv watching

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.

项目摘要 核酸和它们的构建单元在所有细胞事件中起着核心作用，因此具有巨大的生物学功能。 这对疾病的出现，进而对人类健康产生了影响。研究这些事件是复杂的， 天然核碱基的非发射性质，这经常使研究人员无法使用 现代荧光技术。因此，忠实的最小扰动发射核苷替代物可以 促进监测核苷、核苷酸和核酸在核苷/核苷酸上的转化， “决议”，并推进基础研究，诊断工具和药物发现的努力。 该项目的目标是设计和合成新的同晶发射核苷， 核苷酸类似物，并将它们作为探针用于监测基于核苷和核苷酸的 转化以及核酸的功能、结构、动力学和识别。具体来说，少校 将应对当代的挑战，努力弥合主要差距，其中包括： 生物物理技术，如荧光检测圆二色性（FDCD），介绍了近五个 （B）多光子、成像和单分子光谱学- 基于实验，使用天然或最小干扰的寡核苷酸或核苷酸辅因子， （c）类似地，核苷/核苷酸加工酶的单分子酶学没有得到充分利用。 （d）用于基本过程的真实的时间探测的探针，例如肽基转移酶、相位 缺乏分离的无膜细胞器形成和mRNA衰变;（e）基于核苷/潮的 不能直接监测代谢过程和基于核苷酸的信号传导事件;和（f）高 核苷和核苷加工酶的通量筛选不能实时进行 并且以高生产量的方式而不使用可靠的发射替代衬底。 利用我们实验室开发的几个有用的发射性核苷替代物家族， 将进一步完善我们的“设计师”发射和同构核苷/潮汐，并将其应用于推进 解决上述挑战的方法。我们将追求新的物理和 生化方法以及有效的实时筛查和诊断工具。这些努力将扩大 社区的发射功能探针库，推动未来的发现和成像 应用.这些创新反过来将进一步从根本上理解关键的生物过程 这与疾病发展有关，并将对改善人类健康产生长期影响。