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光辅助检测和测序 5-甲酰胞苷 RNA。

Photo-Facilitated Detection and Sequencing of 5-Formylcytidine RNA.

机构信息

Beijing National Laboratory for Molecular Sciences (BNLMS), CAS Key Laboratory of Molecular Recognition and Function, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, China.

China National Center for Bioinformation, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China.

出版信息

Angew Chem Int Ed Engl. 2022 Dec 5;61(49):e202210652. doi: 10.1002/anie.202210652. Epub 2022 Nov 10.

Abstract

5-Formylcytidine (f C) is one of the epigenetic nucleotides in tRNA. Despite the evident importance of f C in gene expression regulation, little is known about its exact amount and position. To capture this information, we developed a modification-specific functionalization with a semi-stabilized ylide. The chemical labelling exhibited a high selectivity towards f C and allowed distinction from similar 5-formyluridine. We realized a detection strategy based on the fluorescence signal of the cyclization product 4,5-pyridin-2-amine-cytidine paC, which exhibited a high quantum yield. The results clearly identified f C with a limit of detection at 0.58 nM. This method altered the hydrogen bonding activities of f C and modulated its reverse transcription signature in its sequencing profile. We showed that f C can be detected from tRNA segments with a single-base resolution. Taken together, this approach is a sensitive, antibody-free, and applicable detection and sequencing method for f C-containing RNA.

摘要

5-甲酰胞苷(fC)是 tRNA 中的一种表观遗传核苷酸。尽管 fC 在基因表达调控中的重要性显而易见,但对于其确切的数量和位置知之甚少。为了获取此信息,我们开发了一种针对 fC 的特异性修饰功能化方法,使用半稳定的叶立德。化学标记对 fC 表现出高选择性,并能够与类似的 5-甲酰尿嘧啶区分开来。我们基于环化产物 4,5-吡啶-2-胺胞苷 paC 的荧光信号实现了一种检测策略,该产物具有高量子产率。结果清楚地表明,fC 的检测限为 0.58 nM。该方法改变了 fC 的氢键活性,并在其测序图谱中调节了其逆转录特征。我们表明,fC 可以从具有单碱基分辨率的 tRNA 片段中检测到。总之,该方法是一种灵敏、无抗体且适用于检测和测序含 fC 的 RNA 的方法。

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