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Visualizing RNA in Live Bacterial Cells Using Fluorophore- and Quencher-Binding Aptamers
Murat Sunbul1, Ankita Arora1, Andres Jäschke2
1Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, Heidelberg, 69120, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|November 14, 2017
Summary
Researchers developed a new live-cell RNA imaging method. This technique uses aptamers and fluorophores to visualize RNA in living cells with minimal disruption.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Visualizing RNA of interest (ROI) in living cells is crucial for understanding RNA roles, dynamics, and regulation.
- Noninvasive imaging methods are needed to study RNA function without perturbing cellular processes.
Purpose of the Study:
- To develop a novel live-cell RNA imaging method for noninvasive visualization of RNA in living cells.
- To enable multicolor and dual-color imaging of distinct RNAs in live bacteria.
Main Methods:
- Genetically fusing fluorophore- and quencher-binding aptamers to the RNA of interest (ROI).
- Utilizing membrane-permeable, nonfluorescent fluorophore-quencher conjugates that become fluorescent upon aptamer binding.
- Employing small RNA tags and small molecule fluorophores for labeling.
Main Results:
- Demonstrated a dramatic increase in fluorescence upon fluorophore-quencher conjugate binding to aptamers.
- Successfully achieved labeling of ROIs with multiple distinct colors.
- Enabled dual-color imaging of two different RNAs simultaneously in live bacteria.
Conclusions:
- The developed method provides a novel approach for live-cell RNA imaging.
- The use of small tags and fluorophores minimizes perturbation to RNA function and dynamics.
- This technique facilitates multicolor and dual-color RNA visualization in live bacteria, aiding in the study of RNA biology.
