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Updated: Oct 4, 2025
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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
11.9K
Development of a split fluorescent protein-based RNA live-cell imaging system to visualize mRNA distribution in
Nien-Chen Huang1, Kai-Ren Luo1, Tien-Shin Yu2
1Institute of Plant and Microbial Biology, Academia Sinica, Taipei, 11529, Taiwan.
Plant Methods
|February 8, 2022
Summary
This study introduces a novel RNA live-cell imaging system that eliminates nuclear background fluorescence. The new method enables clearer visualization of mRNA distribution in living plant cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Biotechnology
Background:
- RNA live-cell imaging visualizes subcellular mRNA distribution using RNA-binding proteins (RBPs).
- Existing RBP-based systems can suffer from nuclear background fluorescence, hindering signal detection.
- Nuclear localization sequences used to reduce background can interfere with visualizing nuclear RNA.
Purpose of the Study:
- To develop an improved RNA live-cell imaging system with reduced background fluorescence.
- To overcome limitations of existing methods for visualizing mRNA in living cells.
Main Methods:
- Utilized a split fluorescent protein (FP)-based approach, fusing RBPs to FP fragments.
- Co-expressed RBPs with target mRNA linked to RNA recognition sequences to reconstitute functional FPs.
- Optimized the system for minimal background fluorescence.
Main Results:
- Successfully eliminated background fluorescence in the nucleus.
- Developed a functional split FP-based RNA imaging system.
- Demonstrated the system's efficacy in visualizing mRNA in living plant cells.
Conclusions:
- Established a background-free RNA live-cell imaging system.
- Provided a new platform for visualizing subcellular mRNA distribution in living plant cells.
