Antibody-Free Fluorine-Assisted Metabolic Sequencing of RNA N 4 -Acetylcytidine.
Shen YanZiang LuWei YangJinglei XuYafen WangWei XiongRongjie ZhuLinao RenZhaoxin ChenQi WeiSong-Mei LiuTian FengBi-Feng YuanXiaocheng WengYuhao DuXiang ZhouPublished in: Journal of the American Chemical Society (2023)
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.