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 核酸及其组成部分在所有细胞事件中都起着核心作用，因此具有巨大的作用 影响疾病的出现以及对人类健康的影响。研究此类事件由 天然核试剂酶的非放电性质，这经常剥夺研究人员的使用 现代荧光技术。因此，忠实的微不足道的发射核代理人可以 促进核外侧，核侧和基于核酸的核横向/潮汐转化的监测 - “解决方案”，并推进基础研究，诊断工具和药物发现工作。 拟议程序的目的是设计和综合新的同构成构核球体和 核苷酸类似物并将其作为监测基于核苷和核苷酸的问题 转化以及核酸的功能，结构，动力学和识别。具体而言，专业 当代挑战将在试图弥合重大差距的尝试中，其中包括：（a）强大的 生物物理技术，例如荧光检测的圆二色性（FDCD），引入了近五个 几十年前，实际上仍然是出乎意料的。 （b）多光子，成像和单分子光谱学 - 使用天然或微薄的寡核苷酸或核苷酸辅助因子的基于基于的实验是严重的 未充分利用； （c）同样，核外侧/潮汐加工酶的单分子酶学尚未 先进的; （d）实时探索基本过程，例如肽基转移酶，相位的探针 缺乏分离的无膜细胞器形成和mRNA衰减。 （e）基于核苷/潮汐 代谢过程和基于核苷酸的信号事件无法直接监测； （f）高 无法实时进行核苷和核苷加工酶的吞吐量筛选 并以高吞吐量的方式不使用忠实的发射替代底物。 利用在我们的实验室中开发的几个有用的发射核代理人家族，我们 将进一步完善我们的“设计师”的发射和同构核核苷/潮汐，并将其应用于 解决上述挑战的解决方案。我们将追求新的身体和 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的武器库，将未来的进步迈进了发现和成像 申请。这些创新反过来将进一步了解关键生物学过程 与疾病的发展有关，将对改善人类健康产生长期影响。