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4‐Thiouridine‐Enhanced Peroxidase‐Generated Biotinylation of RNA
Author(s) -
Huang Jinguo,
Zhao Ruiqi,
Qin Shanshan,
Yang Shixi,
Li Wei,
Mo Jing,
Wang Fang,
Du Yuhao,
Weng Xiaocheng,
Zhou Xiang
Publication year - 2021
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000567
Subject(s) - biotinylation , rna , subcellular localization , biochemistry , chemistry , transcriptome , microbiology and biotechnology , computational biology , biology , gene expression , gene
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 (s 4 U)‐enhanced peroxidase‐generated biotinylation of RNA with high efficiency. The incorporation of s 4 U could introduce additional sites for RNA labeling, enhanced biotinylation was observed on monomer, model oligo RNA and total RNA. Through the s 4 U metabolic approach, the in vivo RNA biotinylation efficiency by peroxidase catalysis was also dramatically increased, which will benefit RNA isolation and study for the spatial transcriptome.