Antibody-Free Fluorine-Assisted Metabolic Sequencing of RNA <i>N</i><sup>4</sup>-Acetylcytidine
Shen Yan, Ziang Lu, Wei Yang, Jinglei Xu, Yafen Wang, Wei Xiong, Rongjie Zhu, Linao Ren, Zhaoxin Chen, Qi Wei, Song‐Mei Liu, Tian Feng, Bi‐Feng Yuan, Xiaocheng Weng, Yuhao Du, Xiang Zhou
Abstract
N 4 -Acetylcytidine (ac 4 C) has been found to affect a variety of cellular and biological processes. For a mechanistic understanding of the roles of ac 4 C in biology and disease, we present an antibody-free, fluorine-assisted metabolic sequencing method to detect RNA ac 4 C, called “FAM-seq”. We successfully applied FAM-seq to profile ac 4 C landscapes in human 293T, HeLa, and MDA cell lines in parallel with the reported acRIP-seq method. By comparison with the classic ac 4 C antibody sequencing method, we found that FAM-seq is a convenient and reliable method for transcriptome-wide mapping of ac 4 C. Because this method holds promise for detecting nascent RNA ac 4 C modifications, we further investigated the role of ac 4 C in regulating chemotherapy drug resistance in chronic myeloid leukemia. The results indicated that drug development or combination therapy could be enhanced by appreciating the key role of ac 4 C modification in cancer therapy.