Protocol for transcriptome-wide mapping of small-molecule RNA-binding sites in live cells

被引：1
|
作者：
Tong, Yuquan [1 ,2 ]
Zanon, Patrick R. A. [1 ]
Yang, Xueyi [1 ,2 ]
Su, Xiaoxuan [1 ,2 ]
Childs-Disney, Jessica L. [1 ]
Disney, Matthew D. [1 ,2 ]
机构：
[1] Herbert Wertheim UF Scripps Inst Biomed Innovat &, Dept Chem, 130 Scripps Way, Jupiter, FL 33458 USA
[2] Scripps Res Inst, Dept Chem, 130 Scripps Way, Jupiter, FL 33458 USA
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 03期
基金：
美国国家卫生研究院;
关键词：
D O I：
10.1016/j.xpro.2024.103271
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Small molecules targeting RNA can be valuable chemical probes and potential therapeutics. The interactions between small molecules, particularly fragments, and RNA, however, can be difficult to detect due to their modest affinities and short residence times. Here, we present a protocol for mapping the molecular fingerprints of small molecules in vitro and throughout the human transcriptome in live cells. We describe steps for compound treatment, cross-linking, RNA extraction, fragmentation, and pull-down. We then detail procedures for RNA sequencing and data analysis. For complete details on the use and execution of this protocol, please refer to Tong et al.1 1
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页数：31
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