荧光指示剂置换分析用于配体-RNA 相互作用。

Fluorescent indicator displacement assay for ligand-RNA interactions.

机构信息

Department of Regulatory Bioorganic Chemistry, The Institute of Scientific and Industrial Research, Osaka University, Ibaraki 567-0047, Japan.

出版信息

J Am Chem Soc. 2010 Mar 24;132(11):3660-1. doi: 10.1021/ja100089u.

DOI:10.1021/ja100089u

PMID:20196592

Abstract

Since the discovery of the RNA functions modulating gene expression, functional RNA has become an attractive drug target. Ligand-RNA interactions have been studied by covalent labeling of either the ligand or RNA with fluorescent dyes or immobilization on the sensing devices. However, it is desirable to investigate the interactions without any chemical modifications in order to keep the native secondary and tertiary structures. Displacement assays satisfy this criterion because neither covalent labeling nor immobilization is necessary for the assay. We here report a fluorescent indicator displacement assay for the investigation of a ligand-RNA interaction that exploits a novel fluorescent indicator, a 2,7-disubstituted 9H-xanthen-9-one derivative. The assay was found to be applicable for screening new ligands binding to RRE RNA from chemical library.

摘要

自发现 RNA 具有调节基因表达的功能以来，功能性 RNA 已成为一个有吸引力的药物靶点。通过将配体或 RNA 用荧光染料共价标记或固定在传感设备上，已经研究了配体-RNA 相互作用。然而，理想的情况是在不进行任何化学修饰的情况下研究相互作用，以保持天然的二级和三级结构。置换测定满足了这一标准，因为测定既不需要共价标记也不需要固定。我们在这里报告了一种荧光指示剂置换测定法，用于研究配体-RNA 相互作用，该方法利用了一种新型的荧光指示剂，即 2,7-二取代的 9H-呫吨-9-酮衍生物。该测定法被发现适用于从化学文库中筛选与 RRE RNA 结合的新配体。