Synthesis of Sensitive Epitranscriptomically Modified RNAs

项目摘要/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。 含有敏感官能团的表位转录修饰的RNA。这里是敏感的官能团 指那些在碱性和亲核条件下不稳定的物质，广泛用于解除保护和 在现有的化学RNA合成技术中的裂解。许多自然产生的RNA包括tRNA， RRNA、mRNA、SnRNA、snoRNA、miRNA和lncRNA都包含这样的敏感基团。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。为了获得诸如敏感群体调节RNA的机制等知识 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，许多这样的RNA不能由任何 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、裂解和去保护 到目前为止，在生命系统中发现的几乎所有敏感群体都是稳定的条件，因此新的 技术将能够合成几乎所有自然产生的敏感RNA。为了说明预测的情况 影响广泛的敏感RNA合成技术，模型中含有敏感的ac4C的mRNAs 将使用该技术合成修饰。Ac4C的异常调节与许多人类 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 In可促进蛋白质合成，但其分子机制尚不清楚。包含ac4C的模型 MRNAs将被用来填补这一知识空白。新的知识预计将在以下领域有用 确定治疗各种人类疾病的新的治疗靶点，并评估 作为药物的含有AC4C的RNA。除了这个具体的例子之外，还可以获得广泛的敏感信息 RNA，许多其他生物医学项目，如识别负责阅读和擦除的蛋白质 各种RNA修饰，使用含有敏感修饰的RNA作为机器的训练样本 了解纳米孔单分子RNA测序和破译敏感基团的机制 调节RNA降解和蛋白质合成，目前所有这些都是不可能或具有挑战性的， 将成为生物医学研究界的可能。

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.