Issue 48, 2023

A universal orthogonal imaging platform for living-cell RNA detection using fluorogenic RNA aptamers

Abstract

MicroRNAs (miRNAs) are crucial regulators of gene expression at the post-transcriptional level, offering valuable insights into disease mechanisms and prospects for targeted therapeutic interventions. Herein, we present a class of miRNA-induced light-up RNA sensors (miLS) that are founded on the toehold mediated principle and employ the fluorogenic RNA aptamers Pepper and Squash as imaging modules. By incorporating a sensor switch to disrupt the stabilizing stem of these aptamers, our design offers enhanced flexibility and convertibility for different target miRNAs and aptamers. These sensors detect multiple miRNA targets (miR-21 and miR-122) with detection limits of 0.48 and 0.2 nM, respectively, while achieving a robust signal-to-noise ratio of up to 44 times. Capitalizing on the distinct fluorescence imaging channels afforded by Pepper-HBC620 (red) and Squash-DFHBI-1T (green), we establish an orthogonal miRNA activation imaging platform, enabling the simultaneous visualization of different intracellular miRNAs in living cells. Our dual-color orthogonal miLS imaging platform provides a powerful tool for sequence-specific miRNA imaging in different cells, opening up new avenues for studying the intricate functions of RNA in living cells.

Graphical abstract: A universal orthogonal imaging platform for living-cell RNA detection using fluorogenic RNA aptamers

Supplementary files

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Article information

Article type
Edge Article
Submitted
20 Sep 2023
Accepted
21 Nov 2023
First published
02 Dec 2023
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2023,14, 14131-14139

A universal orthogonal imaging platform for living-cell RNA detection using fluorogenic RNA aptamers

P. Yin, M. Ge, S. Xie, L. Zhang, S. Kuang and Z. Nie, Chem. Sci., 2023, 14, 14131 DOI: 10.1039/D3SC04957D

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