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.

项目概要 核酸及其构建模块在所有细胞事件中发挥着核心作用，因此具有巨大的作用。 影响疾病的出现，进而影响人类健康。研究此类事件很复杂 天然核碱基的非发射性质，这常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 促进在核苷/潮汐条件下监测核苷、核苷酸和核酸的转化 “决议”，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构发射核苷和 核苷酸类似物并将其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，主要 将解决当代挑战，试图弥合重大差距，其中包括： (a) 强大 生物物理技术，例如荧光检测圆二色性（FDCD），引入了近五种 几十年前，实际上仍未被探索过； (b) 多光子、成像和单分子光谱学- 使用天然或最小干扰的寡核苷酸或核苷酸辅因子的基于实验，严重 未充分利用； (c) 同样，核苷/潮汐加工酶的单分子酶学还没有 先进的; (d) 实时探索肽基转移酶、相等基本过程的探针 缺乏分离的无膜细胞器形成和 mRNA 降解； (e) 核苷/潮汐基 代谢过程和基于核苷酸的信号传导事件无法直接监测； (f) 高 无法实时进行核苷和核苷加工酶的通量筛选 并以高通量方式进行，无需使用忠实的发射替代基板。 利用我们实验室开发的几个有用的发射核苷替代物家族，我们 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 针对上述挑战的解决方案。我们将追求新物理和新材料的进步 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，推动未来在发现和成像领域的进步 应用程序。这些创新反过来将进一步加深对关键生物过程的基本理解 与疾病的发展相关，将对改善人类健康产生长期影响。