Determination of RNA-ligand interactions with the photoaffinity platform PEARL-seq.
Determination of RNA-ligand interactions with the photoaffinity platform PEARL-seq.
复制标题
使用光亲和平台 PEARL-seq 测定 RNA-配体相互作用。
DOI：
10.1016/j.ymeth.2022.06.009
复制
发表时间：
2022
期刊：
影响因子：
4.8
通讯作者：
J. Craig Blain
中科院分区：
文献类型：
--
作者：
A. N. Sexton;Lee E. Vandivier;Jennifer C Petter;Herschel Mukherjee;J. Craig Blain
在治疗剂的开发中，重要的是建立化合物与其预期靶标的接合并鉴定其结合的其他靶标。因此，需要用于在RNA靶向治疗的不断增长的领域中证明靶标接合的方法。我们提出了一个详细的方案，用于RNA连接测序（PEARL-seq）的光亲和性评估，这是一个用于确定小分子配体与其靶RNA之间相互作用的平台。PEARL-seq允许以单核苷酸分辨率检测结合和交联事件，并允许测量靶RNA相对于所有其他RNA的富集。PEARL-seq是验证真正的RNA-配体相互作用的有价值的工具。
In the development of therapeutics, it is important to establish engagement of a compound to its intended target and identify other targets it binds to. Methods for demonstrating target engagement in the growing field of RNA-targeted therapeutics are therefore needed. We present a detailed protocol for Photoaffinity Evaluation of RNA Ligation-Sequencing (PEARL-seq), a platform for determining interactions between small molecule ligands and their target RNA(s). PEARL-seq allows detection of binding and crosslinking events with single nucleotide resolution and allows measurement of enrichment of the target RNA relative to all other RNAs. PEARL-seq is a valuable tool in the effort to verify bona fide RNA-ligand interactions.