RNA-mediated activity-based protein profiling reveals the substrates of a human dihydrouridine synthase

We adapted the iCLIP method to map DUS3L-dependent DHU sites in a transcriptome-wide manner by treating the cells with 5-FUrd. Overall design: Flp-In T-Rex 293 cells expressing 3xFlag-DUS3L were treated with 5-FUrd, DUS3L-RNA were extracted and subject for Iclip library preparation.

本研究通过以5-氟尿苷（5-FUrd）处理细胞，对紫外交联免疫沉淀测序（iCLIP）方法进行适配改造，以在全转录组范围内绘制依赖于DUS3L的二氢尿嘧啶（DHU）位点。整体实验设计：将表达3xFlag-DUS3L的Flp-In T-Rex 293细胞用5-FUrd处理，提取结合DUS3L的RNA，并进行iCLIP文库制备。