Department of Chemistry & Pharmaceutical Sciences, VU University Amsterdam, De Boelelaan 1108, 1081, HZ, Amsterdam, The Netherlands.
Angew Chem Int Ed Engl. 2019 Jan 28;58(5):1266-1279. doi: 10.1002/anie.201806482. Epub 2018 Nov 5.
The cellular functions of RNA are not limited to their role as blueprints for protein synthesis. In particular, noncoding RNA, such as, snRNAs, lncRNAs, miRNAs, play important roles. With increasing numbers of RNAs being identified, it is well known that the transcriptome outnumbers the proteome by far. This emphasizes the great importance of functional RNA characterization and the need to further develop tools for these investigations, many of which are still in their infancy. Fluorescent light-up aptamers (FLAPs) are RNA sequences that can bind nontoxic, cell-permeable small-molecule fluorogens and enhance their fluorescence over many orders of magnitude upon binding. FLAPs can be encoded on the DNA level using standard molecular biology tools and are subsequently transcribed into RNA by the cellular machinery, so that they can be used as fluorescent RNA tags (FLAP-tags). In this Minireview, we give a brief overview of the fluorogens that have been developed and their binding RNA aptamers, with a special focus on published crystal structures. A summary of current and future cellular FLAP applications with an emphasis on the study of RNA-RNA and RNA-protein interactions using split-FLAP and Förster resonance energy transfer (FRET) systems is given.
RNA 的细胞功能不仅限于作为蛋白质合成蓝图的作用。特别是非编码 RNA，如 snRNA、lncRNA、miRNA，发挥着重要作用。随着越来越多的 RNA 被鉴定出来，人们已经清楚地认识到转录组远远超过了蛋白质组。这强调了对功能性 RNA 特征进行描述的重要性，并需要进一步开发这些研究的工具，其中许多仍处于起步阶段。荧光点亮适体（FLAP）是可以结合非毒性、细胞可渗透的小分子荧光团的 RNA 序列，并在结合后将其荧光增强许多数量级。FLAP 可以使用标准的分子生物学工具在 DNA 水平上进行编码，然后由细胞机制转录成 RNA，因此可以用作荧光 RNA 标签（FLAP-tag）。在这篇综述中，我们简要概述了已经开发的荧光团及其结合的 RNA 适体，特别关注已发表的晶体结构。总结了当前和未来的细胞 FLAP 应用，重点介绍了使用分裂 FLAP 和Förster 共振能量转移（FRET）系统研究 RNA-RNA 和 RNA-蛋白质相互作用。
