基于化学标记的单碱基分辨率绘制转录组中5-醛基胞嘧啶谱图的研究

RNA修饰是转录后调控研究的热点，这些修饰与疾病的发生密切相关，为了研究RNA修饰碱基在转录组中的分布规律，以探讨它们的生物学功能以及和疾病产生的关系，需要开发更多绘制RNA修饰碱基的单碱基分辨率谱图的技术。本项目拟基于化学生物学的思路和手段，寻找能识别RNA中5-醛基胞嘧啶的分子探针，在单碱基分辨率水平上绘制转录组中5-醛基胞嘧啶的谱图。主要研究思路为，通过小分子的设计和功能评价，筛选最优分子探针，使标记后的5-醛基胞嘧有效富集。通过逆转录，标记的5-醛基胞嘧啶易于与腺嘌呤配对，具体体现在测序结果读作胸腺嘧啶，结合高通量测序，实现转录组中5-醛基胞嘧啶单碱基分辨率谱图的绘制。该研究工作将有助于扩展RNA修饰核酸研究领域，为探索修饰核酸的生物医学功能中的作用提供研究基础。

The study of RNA modification has been one of the research focuses in post-transcriptional regulation. RNA modifications are closely related to the occurrence of diseases. In order to study the distribution of RNA-modified nucleobases in the transcriptome, and to discuss their biological functions and the relationship with diseases, more techniques to profile single-base resolution maps of RNA-modified bases need to be developed. Based on chemical biology principle, this project intends to search for molecular probes that can identify 5-formylcytidine (f5C) in RNA, and profile f5C in the human transcriptome at the single-base resolution level. The main research idea is to design and select the optimal molecular probe for effectively enriching the labeled 5-formylcytidine. Through reverse transcription, labelling f5C enabled C-to-T signals in the sequencing reads. Such C-to-T transition of f5C was used from the sequencing reads to obtain transcriptome-wide maps of f5C at single-nucleotide resolution. This work will expand the field of RNA modified nucleic acid research and provide a research basis for exploring the biological functions of modified nucleic acid.