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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

Near-infrared fluorogenic RNA for in vivo imaging and sensing.

Zhenyin Chen^1,2,3, Wei Chen^4,5, Cun Xu^6,7

¹Interdisciplinary Science Center, State Key Laboratory of Integrated Management of Pest Insects and Rodents,Institute of Zoology, Chinese Academy of Sciences, 100101, Beijing, China.

Nature Communications

|January 9, 2025

Summary

Researchers developed a novel near-infrared (NIR) fluorescent RNA tag using a fluorogenic RNA called Squash and a fluorophore DFQL-1T. This advancement enables RNA visualization and sensing in living cells and animals for in vivo imaging applications.

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Area of Science:

Molecular Biology
Biochemistry
Biotechnology

Background:

Fluorogenic RNA aptamers are used for RNA imaging and sensing in cells.
Current RNA:fluorophore complexes emit visible light, limiting in vivo applications.
Developing near-infrared (NIR) fluorescent RNA tags for in vivo studies remains a challenge.

Purpose of the Study:

To create a novel fluorogenic RNA:fluorophore complex emitting near-infrared (NIR) fluorescence.
To enable RNA visualization and sensing in living mammalian cells and mice.
To develop RNA-based sensors for detecting specific targets in vivo.

Main Methods:

Identification and modulation of red fluorescent protein-like fluorophores for binding to the Squash RNA aptamer.
Characterization of the photophysical properties of the Squash:DFQL-1T complex.
Application of the complex for RNA visualization in mammalian cells and mice.
Development of RNA-based sensors for non-coding RNAs and small molecules.

Main Results:

A novel fluorogenic RNA:fluorophore complex, Squash:DFQL-1T, was identified.
The complex exhibits photostable near-infrared (NIR) fluorescence.
Successful visualization of RNA in living mammalian cells and mice.
Demonstrated utility of RNA-based sensors for target detection in vivo.

Conclusions:

The Squash:DFQL-1T complex represents a breakthrough in developing NIR fluorescent RNA tags.
This technology facilitates in vivo RNA imaging and sensing in biological systems.
The developed complex can be adapted for diverse applications in molecular biology and medicine.