Fluorescent Nucleosides and Oligonucleotides

荧光核苷和寡核苷酸

• 批准号: 8845949
• 负责人: YITZHAK TOR
• 金额: $ 5.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8845949
• 关键词: Address; Antibiotics; Behavior; Biochemical; Biological Assay; Biological Process; Biology; Brain; Cell Nucleus; Characteristics; Chemicals; Complement; DNA; Data; Deamination; Development; Diagnostic; Disease; Event; Family; Fluorescence; Generations; Genetic; Goals; Health; Human; Investigation; Knowledge; Ligands; Link; Messenger RNA; Metabolism; Methods; Monitor; Motor; Nature; Nucleic Acids; Nucleosides; Nucleotides; Oligonucleotides; Pathway interactions; Phenotype; Play; Predisposition; Process; Property; Protein Biosynthesis; Proteins; Pyrimidine; RNA; RNA Helicase; RNA Interference; Ribosomal Frameshifting; Ribosomes; Role; Shapes; Structure; Techniques; Therapeutic Agents; Time; Virus; absorption; analog; base; design; drug discovery; improved; new therapeutic target; novel; novel diagnostics; novel strategies; nucleic acid structure; nucleobase; nucleoside analog; programs; protein expression; quantum; tool; trend; tv watching

DESCRIPTION (provided by applicant): The goal of the proposed program is to design and synthesize new fluorescent nucleoside analogs and implement them as probes for nucleic acids structure, dynamics and recognition. Advancing effective fluorescence-based tools for exploring nucleic acids and their interactions with ligands and potential therapeutic agents will further new diagnostic approaches and will facilitate drug discovery. The specific aims of this project are: AIM 1. To design, synthesize and incorporate new isomorphic fluorescent nucleoside analogs. The main design criteria include: (i) High structural similarity to the native nucleobases to faithfully mimic their size and shape, as well as hybridization and recognition properties, (ii) Re shifted absorption spectrum to minimize overlap with the absorption of the natural bases, and (iii) Adequate emission quantum efficiency and long emission wavelengths (preferably in the visible range). Efficient synthetic pathways will be devised, providing the nucleosides and the necessary building blocks for automated and enzymatic oligonucleotide synthesis. AIM 2. To photophysically and biophysically characterize the modified nucleosides and oligonucleotides. The photophysical characteristics (e.g., absorption and emission maxima, quantum yield and brightness, excited state lifetime, as well as susceptibility to environmental polarity and static and dynamic quenching by native nucleosides) will be rigorously evaluated and interpreted. AIM 3. To implement the promising emissive analogs in biophysical and discovery assays. These assays will facilitate: (i) The discovery of new antibiotics targeting the bacterial ribosome (ii) The study of RNA helicases, ubiquitous motor proteins, which are involved in nearly every aspect of RNA metabolism, (iii) The monitoring of programmed ribosomal frameshifting, a processes which could be extremely detrimental to native protein synthesis, but, when programmed (e.g., in viruses) can maximize protein expression, (iv) The study of RNA deamination, an important posttranscriptional process, which diversifies mRNAs and the resultant proteins; it is linked to proper brain function and, when defective, to disease, and (v) The study of RNAi, a regulatory process induced by short interfering RNA (siRNA), which is also a powerful tool capable of altering cellular phenotypes, deciphering genetic pathways and identifying new therapeutic targets. Nucleic acids play central roles in cellular events and, as such, have immense impact on the emergence of diseases and, in turn, on human health. This necessitates the development of new effective tools for studying their recognition properties and alteration by exogenous agents. The emissive nucleoside analogs designed and prepared will be implemented in novel real time fluorescence-based assays. These investigations will further the fundamental understanding of key biological processes related to disease development and will have long-term impact on improving human health by advancing knowledge and facilitating drug discovery.

描述（由申请人提供）：拟议程序的目标是设计和合成新的荧光核苷类似物，并将其作为核酸结构，动力学和识别的探针实现。推进有效的基于荧光的工具，用于探索核酸及其与配体和潜在治疗剂的相互作用将进一步新 诊断方法并将促进药物发现。该项目的具体目的是：目标1。设计，合成并结合新的同构荧光核苷类似物。主要的设计标准包括：（i）与天然核仁酶的高结构相似性，以忠实地模仿其大小和形状，以及杂交和识别特性，（ii）重新移动吸收光谱，以最大程度地减少与自然基础的吸收的重叠，以及（iii）（iii）适度的发射量子量的效率和长度（偏见），以相互效率和长度（偏爱），以相比（偏见），以范围内（均构成范围内）。将设计有效的合成途径，为自动化和酶促寡核苷酸合成提供核苷和必要的构件。 目的2。以光体物理和生物物理的特征来表征修饰的核苷和寡核苷酸。光物理特征（例如，吸收和发射最大值，量子产量和亮度，激发态寿命，以及对环境极性的敏感性以及对天然核苷的静态和动态淬火）将得到严格评估和解释。 目的3。在生物物理和发现分析中实施有希望的发射类似物。 These assays will facilitate: (i) The discovery of new antibiotics targeting the bacterial ribosome (ii) The study of RNA helicases, ubiquitous motor proteins, which are involved in nearly every aspect of RNA metabolism, (iii) The monitoring of programmed ribosomal frameshifting, a processes which could be extremely detrimental to native protein synthesis, but, when programmed (e.g., in viruses)可以最大程度地提高蛋白质表达，（iv）RNA脱氨基的研究，RNA脱氨基是一个重要的转录后过程，它使mRNA和所得蛋白质多样化；它与适当的大脑功能以及有缺陷的疾病有关，以及（v）RNAi的研究，RNAi是由短干扰RNA（siRNA）引起的调节过程，该过程也是一种能够改变细胞表型，解密遗传途径并识别新的治疗靶标的强大工具。 核酸在细胞事件中起着核心作用，因此对疾病的出现产生了巨大影响，进而对人类健康产生了巨大影响。这需要开发新的有效工具来研究其识别特性和外源性剂的改变。设计和制备的发射核苷类似物将在基于新型的实时荧光测定中实现。这些调查将进一步了解与疾病发展有关的关键生物学过程的基本理解，并通过推进知识和促进药物发现对改善人类健康产生长期影响。