A ratiometric fluorescence RRE RNA-targeted assay for a new fluorescence ligand
A ratiometric fluorescence RRE RNA-targeted assay for a new fluorescence ligand
复制标题
新荧光配体的比率荧光 RRE RNA 靶向测定
DOI：
10.1016/j.bios.2016.06.051
复制
发表时间：
2016-12-15
影响因子：
12.6
通讯作者：
Zhang, Zhi-Qi
中科院分区：
文献类型：
--
作者：
Qi, Liang;Wei, Jing-Ru;Zhang, Zhi-Qi
The Rev protein regulates HIV-1 gene expression. Small ligands that bind to the Rev response element (RRE) RNA would inhibit Rev function and suppress HIV-1 replication. A novel ratiometric fluorescence assay was applied in the present study to monitor ligand-RNA interactions by using red-emitting CdTe quantum dots (QDs) coated with silica as a reference. A small fluorescence ligand, ICR 191, was found to interact with RRE at the Rev binding site and compete with the Rev peptide. After adding red-emitting QDs to the interaction system, it was observed that ICR 191 did not fluoresce upon the addition of RRE, and fluorescence recovered when ICR 191 was displaced by a Rev model peptide, whereas the fluorescence of QDs remained constant. Furthermore, variations in the fluorescence ratios between ICR 191 and QDs were exploited to characterize the interactions of Rev with two known antagonists, neomycin B and tobramycin, by using RRE RNA with ICR 191 as a fluorescence indicator. Together, our results demonstrated that ratiometric fluorescence-based nanotechnology applications can be used for ligand RNA interaction assays. This ICR 191-RRE RNA interaction assay can potentially be developed to build a screening model for assessing antagonists of the Rev binding element in RRE. (C) 2016 Elsevier B.V. All rights reserved.
