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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

Ratiometric sandwich-type assays for RNAs with a point mutation using benzo[a]pyrene-modified probes.

Yu Watari¹, Kaito Nakatani¹, Kentaro Kobata¹

¹Faculty of Molecular Chemistry and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, 606-8585, Japan.

Chemical Communications (Cambridge, England)

|July 3, 2024

Summary

New probes detect RNA point mutations using distinct fluorescence signals. This allows for accurate differentiation between matched and mismatched RNA sequences.

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Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

Published on: August 14, 2018

Area of Science:

Biochemistry
Molecular Biology
Nucleic Acid Chemistry

Background:

Point mutations in RNA are critical biomarkers for various diseases.
Accurate detection of single nucleotide differences in RNA is essential for diagnostics.
Existing methods may lack specificity or require complex procedures.

Purpose of the Study:

To develop novel oligonucleotide probes for sensitive RNA point mutation detection.
To enable discrimination between perfectly matched and single-base mismatched RNA sequences.
To establish a colorimetric and ratiometric detection system.

Main Methods:

Synthesis of Benzo[a]pyrene-modified oligonucleotides.
Hybridization assays with target RNA sequences containing point mutations.
Fluorescence spectroscopy for signal detection and analysis.
Colorimetric and ratiometric measurements.

Main Results:

The developed probes generated two distinct fluorescence signals.
These signals effectively differentiated between matched and single-base mismatched RNA sequences.
Successful colorimetric and ratiometric discrimination was achieved.

Conclusions:

Benzo[a]pyrene-modified oligonucleotides offer a sensitive and specific method for RNA point mutation detection.
The dual-signal fluorescence system provides a robust platform for molecular diagnostics.
This approach facilitates accurate identification of single nucleotide variations in RNA.