4-Thiouridine-Enhanced Peroxidase-Generated Biotinylation of RNA.
4-Thiouridine-Enhanced Peroxidase-Generated Biotinylation of RNA.
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4-硫尿苷增强的过氧化物酶产生的 RNA 生物素化。
DOI：
10.1002/cbic.202000567
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发表时间：
2020
期刊：
影响因子：
3.2
通讯作者：
Zhou Xiang
中科院分区：
文献类型：
--
作者：
Huang Jinguo;Zhao Ruiqi;Qin Shanshan;Yang Shixi;Li Wei;Mo Jing;Wang Fang;Du Yuhao;Weng Xiaocheng;Zhou Xiang
过氧化物酶邻近标记被广泛用于研究活细胞中的亚细胞蛋白质组和蛋白质相互作用网络，但亚细胞RNA标记技术的发展受到限制。APEX-SEQ已成为研究活细胞中亚细胞RNA的一种新方法，但对RNA的标记仍有改进的空间。在这项工作中，我们描述了4-硫代尿苷(S4U)促进的过氧化物酶生物素化的RNA高效。S4U的掺入可以为RNA标记引入额外的位点，单体、模型寡聚RNA和总RNA上的生物素化作用增强。通过S4U代谢途径，大大提高了过氧化物酶催化 活体核糖核糖核酸的生物素化效率，为空间转录组学研究奠定了基础。
Peroxidase‐generated proximity labeling is in widespread use to study subcellular proteomes and the protein interaction networks in living cells, but the development of subcellular RNA labeling is limited. APEX‐seq has emerged as a new method to study subcellular RNA in living cells, but the labeling of RNA still has room to improve. In this work, we describe 4‐thiouridine (s4U)‐enhanced peroxidase‐generated biotinylation of RNA with high efficiency. The incorporation of s4U could introduce additional sites for RNA labeling, enhanced biotinylation was observed on monomer, model oligo RNA and total RNA. Through the s4U metabolic approach, thein vivoRNA biotinylation efficiency by peroxidase catalysis was also dramatically increased, which will benefit RNA isolation and study for the spatial transcriptome.