Synthesis of Sensitive Epitranscriptomically Modified RNAs

敏感表观转录组修饰 RNA 的合成

基本信息

批准号：
10730262
负责人：
Shiyue Fang
金额：
$ 45.15万
依托单位：
MICHIGAN TECHNOLOGICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-02-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10730262
关键词：
Address Affect Area Biological Biological Process Biomedical Research Chemicals Communities Data Development Diabetes Mellitus Disease Enzymes Evaluation Excision Foundations Goals Knowledge Link Machine Learning Medicine Messenger RNA MicroRNAs Modeling Modification Molecular Nature Neurodegenerative Disorders Nucleic Acids Nucleosides Obesity Organism Pathology Phase Prevention Property Protein Biosynthesis Proteins RNA RNA Degradation RNA Stability RNA chemical synthesis RNA-Protein Interaction Reading Research Personnel Research Project Grants Ribosomal RNA Role S phase Sampling Small Nuclear RNA Small Nucleolar RNA Solid Structure Sulfur Technology Thermodynamics Time Training Transfer RNA Translations Writing acyl group amino group base biophysical properties chemical synthesis diagnostic tool disease diagnosis epitranscriptomics functional group human disease innovation leukemia monomer nanopore new technology new therapeutic target nucleobase oxidation single molecule success tool transcriptome sequencing transcriptomics

项目摘要

Project Summary/Abstract The objective of this project is to develop a solid phase technology that is universally useful for the synthesis of epitranscriptomically modified RNAs containing sensitive functional groups. Sensitive functional groups here refers to those that are unstable under the basic and nucleophilic conditions widely used for deprotection and cleavage in existing chemical RNA synthesis technologies. Many naturally occurring RNAs including tRNA, rRNA, mRNA, snRNA, snoRNA, miRNA and lncRNA contain such sensitive groups. They have important biological functions, and errors related to their installation, reading and erasing are associated with human diseases. To obtain the knowledge such as the mechanisms by which the sensitive groups modulate RNA biophysical properties and RNA-protein interactions, and the pathology of diseases involving sensitive groups, chemical synthesis of the sensitive RNAs are needed. Currently many such RNAs cannot be synthesized by any existing technologies. The objective of this project is to fill this technology gap. Using a new set of solid phase synthesis linkers and protecting groups, RNAs will be synthesized, cleaved and deprotected under mild conditions under which almost all sensitive groups found in living systems so far are stable, and thus the new technology will be capable to synthesize almost all naturally occurring sensitive RNAs. To illustrate the predicted broad impact of the sensitive RNA synthesis technology, model mRNAs containing the sensitive ac4C modification will be synthesized using the technology. Dysregulations of ac4C have been linked to many human diseases including leukemia, diabetes, obesity and neurodegenerative diseases. Recent studies found that ac4C in mRNA enhances protein synthesis, but the molecular mechanism is unknown. The ac4C-containing model mRNAs will be used to fill this knowledge gap. The new knowledge is expected to be useful in areas such as identification of new therapeutics targets for treating various human diseases, and evaluation of the potential of ac4C-containing RNAs as medicines. Beyond this specific example, with the access to a wide range of sensitive RNAs, many other biomedical projects such as the identification of proteins responsible for reading and erasing various RNA modifications, the use of RNAs containing sensitive modification as training samples for machine learning in nanopore single molecule RNA sequencing, and deciphering mechanisms by which sensitive groups regulate RNA degradation and protein synthesis, all of which are impossible or challenging to do at this time, will become possible for the biomedical research community.

项目概要/摘要该项目的目标是开发一种普遍适用于合成的固相技术含有敏感功能基团的表观转录修饰RNA。此处敏感官能团指在碱性和亲核条件下不稳定的那些广泛用于脱保护和现有化学RNA合成技术的裂解。许多天然存在的 RNA，包括 tRNA、 rRNA、mRNA、snRNA、snoRNA、miRNA 和 lncRNA 含有此类敏感基团。他们有重要的生物功能以及与其安装、读取和擦除相关的错误与人类有关疾病。获得敏感群体调节RNA机制等知识生物物理特性和RNA-蛋白质相互作用，以及涉及敏感群体的疾病的病理学，需要化学合成敏感的RNA。目前，许多此类 RNA 无法通过任何方法合成。现有技术。该项目的目标是填补这一技术空白。使用一套新的固相合成接头和保护基团，RNA将在温和的条件下合成、切割和脱保护在这些条件下，迄今为止在生命系统中发现的几乎所有敏感群体都是稳定的，因此新的技术将能够合成几乎所有天然存在的敏感RNA。为了说明预测的敏感RNA合成技术的广泛影响，含有敏感ac4C的模型mRNA 将使用该技术合成修饰。 ac4C 的失调与许多人类疾病有关疾病包括白血病、糖尿病、肥胖症和神经退行性疾病。最近的研究发现ac4C mRNA 增强蛋白质合成，但分子机制尚不清楚。包含 ac4C 的模型 mRNA 将用于填补这一知识空白。新知识预计将在以下领域发挥作用：识别治疗各种人类疾病的新治疗靶点，并评估其潜力含有 ac4C 的 RNA 作为药物。除了这个具体的例子之外，还可以访问广泛的敏感信息 RNA，许多其他生物医学项目，例如负责读取和擦除的蛋白质的鉴定各种RNA修饰，使用含有敏感修饰的RNA作为机器的训练样本学习纳米孔单分子 RNA 测序，并破译敏感群体的机制调节RNA降解和蛋白质合成，所有这些目前都是不可能或具有挑战性的，生物医学研究界将成为可能。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

dM-Dim for Carboxylic Acid Protection.

dM-Dim 用于羧酸保护。

DOI：
发表时间：
2018-05-02
期刊：
影响因子：
1.8
作者：
Shahsavari, Shahien;Wigstrom, Travis;Gooding, James;McNamara, Chase;Fang, Shiyue
通讯作者：
Fang, Shiyue

Determination of optical density (OD) of oligodeoxynucleotide from HPLC peak area.

从 HPLC 峰面积测定寡脱氧核苷酸的光密度 (OD)。

DOI：
发表时间：
2022
期刊：
影响因子：
0
作者：
Chillar, Komal;Yin, Yipeng;Eriyagama, Adikari M Dhananjani N;Fang, Shiyue
通讯作者：
Fang, Shiyue

An amine protecting group deprotectable under nearly neutral oxidative conditions.

在近中性氧化条件下可脱保护的胺保护基。

DOI：
发表时间：
2018
期刊：
影响因子：
0
作者：
Shahsavari, Shahien;McNamara, Chase;Sylvester, Mark;Bromley, Emily;Joslin, Savannah;Lu, Bao;Fang, Shiyue
通讯作者：
Fang, Shiyue

Incorporation of Sensitive Ester and Chloropurine Groups into Oligodeoxynucleotides through Solid Phase Synthesis.

通过固相合成将敏感酯和氯嘌呤基团掺入寡脱氧核苷酸中。

DOI：
10.1002/slct.201801484
发表时间：
2018-08-16
期刊：
ChemistrySelect
影响因子：
2.1
作者：
Bhaskar Halami;Shahien Shahsavari;Zack Nelson;Lucas Prehoda;Dhananjani N A M Eriyagama;Shiyue Fang
通讯作者：
Shiyue Fang

Tritylation of Alcohols under Mild Conditions without Using Silver Salts.

在温和条件下不使用银盐对醇进行三苯甲基化。