Fluorescent Nucleosides and Oligonucleotides

荧光核苷和寡核苷酸

基本信息

批准号：
9908107
负责人：
YITZHAK TOR
金额：
$ 38.04万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-01-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9908107
关键词：
Address Adenosine Anticodon Behavior Binding Biochemical Biological Biological Assay Biological Process Biology Biophysics Categories Cell Nucleus Characteristics Chemicals Codon Nucleotides Coenzymes Complement DNA Data Deamination Development Diagnostic Disease Event Family Fluorescence Generations Goals Health Human Innate Immune Response Inosine Investigation Kinetics Knowledge Messenger RNA Metabolism Methods Modification Monitor Nature Nucleic Acid Probes Nucleic Acids Nucleosides Nucleotides Oligonucleotides Outcome Pathway interactions Peptide Synthesis Play Predisposition Process Property Protein Biosynthesis Proteins Pyrimidine RNA RNA Folding RNA-Binding Proteins Ribosomal Frameshifting Ribosomes Role Screening procedure Second Messenger Systems Shapes Structure Therapeutic Agents Time Transfer RNA Translations Virus Replication absorption analog base design drug discovery improved novel novel diagnostics nucleobase nucleoside analog nucleotide analog programs protein expression purine metabolism quantum tool tv watching

项目摘要

PROJECT SUMMARY The goal of the proposed program is to design and synthesize new 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. Advancing effective fluorescence-based tools for exploring nucleoside, nucleotide and oligonucleotides, as well as their metabolism, regulatory processes and interactions with potential therapeutic agents will further knowledge and advance new diagnostic approaches, facilitating drug discovery. Specifically, we will address: AIM 1. To design, synthesize and incorporate new isomorphic fluorescent nucleoside and nucleotides. 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) Red shifted absorption spectrum to minimize overlap with the absorption of the natural bases, and (iii) Adequate emission quantum efficiency and visible emission wavelengths. Efficient synthetic and enzymatic pathways will be devised, providing the nucleosides, nucleotides and oligonucleotide. 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. The outcome of these analyses and the data generated dictate the utility and potential applications of the nucleoside surrogates. AIM 3. To implement the promising emissive analogs in biophysical, biochemical and discovery assays. These assays will facilitate: (i) Studying the enzymatic deamination of adenosine to inosine, which occurs in three key biological contexts: purine metabolism, mRNA editing and tRNA maturation; (ii) Investigating the enzymatic synthesis of emissive second messengers, such as c-di-NMPs, and their interactions with riboswitches and proteins (e.g., STING), (iii) Monitoring RNA–protein binding (e.g., k- turn/7LAe) and translational events, including programmed ribosomal frameshifting, which could be detrimental to native protein synthesis, but, when programmed (e.g., in viral replication) can maximize protein expression. 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.

项目摘要拟议程序的目的是设计和综合新的发射核外侧和核横向类似物并将它们作为监测基于核犹太的转变的问题推进有效的基于荧光的有效探索核外侧，核外侧和寡核苷酸的工具及其新陈代谢，调节性过程和与潜在治疗剂的互动将进一步了解并推进新的知识诊断方法，支持药物发现。具体来说，我们将解决：目标1。设计，合成并结合新的同构荧光核苷和核苷酸。主要的设计标准包括：（i）与天然核基地的高结构相似性忠实地模仿它们的大小和形状，以及杂交和识别特性，（ii）红色抽象光谱以最大程度地减少与自然碱的抽象重叠，（iii）足够的发射量子效率和可见发射波长。有效的合成和酶促途径将是设计，提供核外侧，核外侧和寡核苷酸。目的2。以光体物理和生物物理的特征来表征修饰的核酶和寡核苷酸。光物理特征（例如抽象和发射最大值，量子产率和亮度，激发态寿命以及对环境极性的敏感性以及静态和动态由本地核酶进行淬火）将经过严格评估和解释。这些分析的结果产生的数据决定了核球体替代物的效用和潜在应用。目的3。在生物物理，生化和发现中实施承诺的发射类似物测定。这些评估将有助于：（i）研究腺苷对肌苷的酶促脱氨酸，这发生在三个关键的生物学环境中：纯代谢，mRNA编辑和tRNA成熟；（ii）研究发射第二使者（例如C-DI-NMP及其）的酶促合成与核糖开关和蛋白质（例如，刺）的相互作用（iii）监测RNA - 蛋白质结合（例如，K-， Turn/7lae）和翻译事件，包括编程的核糖体框架，这可能有害对于天然蛋白质的合成，但是，当对其进行编程时（例如，在病毒复制中）可以最大化蛋白质表达。核酸在细胞事件中起着核心作用，因此对疾病，反过来涉及人类健康。这需要开发研究的新工具它们的识别特性和外源性剂的改变。设计的发射核可类似物设计并准备在基于新型的实时荧光测定中实施。这些调查会进一步，对与疾病发展有关的关键生物过程的基本了解，将通过促进知识并支持药物发现对改善人类健康产生长期影响。

项目成果

期刊论文数量（67）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Chemical Mutagenesis of an Emissive RNA Alphabet.

DOI：
10.1021/jacs.5b10420
发表时间：
2015-11-25
期刊：
Journal of the American Chemical Society
影响因子：
15
作者：
Rovira AR;Fin A;Tor Y
通讯作者：
Tor Y

Real-Time Monitoring of Human Guanine Deaminase Activity by an Emissive Guanine Analog.

DOI：
10.1021/acschembio.1c00232
发表时间：
2021-07-16
期刊：
ACS CHEMICAL BIOLOGY
影响因子：
4
作者：
Bucardo, Marcela S.;Wu, You;Ludford, Paul T.;Li, Yao;Fin, Andrea;Tor, Yitzhak
通讯作者：
Tor, Yitzhak

Modified 6-aza uridines: highly emissive pH-sensitive fluorescent nucleosides.

DOI：
10.1002/cphc.201200375
发表时间：
2012-10-08
期刊：
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子：
0
作者：
Sinkeldam RW;Hopkins PA;Tor Y
通讯作者：
Tor Y

Fluorescent ribonucleoside as a FRET acceptor for tryptophan in native proteins.