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Related Experiment Video

Updated: Oct 15, 2025

Fluorescent Visualization of Mango-tagged RNA in Polyacrylamide Gels via a Poststaining Method
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Single-Molecule RNA Imaging Using Mango II Arrays.

Adam D Cawte1,2,3, Haruki Iino1,2, Peter J Unrau4

  • 1Single Molecule Imaging Group, MRC London Institute of Medical Sciences, London, UK.

Methods in Molecular Biology (Clifton, N.J.)
|October 25, 2021
PubMed
Summary

Novel RNA Mango variants offer improved fluorescence and stability for background-free cellular RNA imaging. This protocol details imaging Mango II tagged RNAs in fixed and live cells with single-molecule sensitivity.

Keywords:
Fluorogenic RNA AptamerLive cell imagingRNA dynamicsRNA-MangoSingle-Molecule MicroscopySuperresolution microscopy

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • Fluorogenic RNA aptamers like Spinach and Mango are used for cellular RNA labeling.
  • Previous aptamers had limitations in folding efficiency and fluorescent stability.
  • New RNA-Mango variants show enhanced fluorescence and stability.

Purpose of the Study:

  • To present a protocol for imaging RNA using novel RNA-Mango variants.
  • To enable sensitive detection of cellular RNAs in both fixed and live cells.

Main Methods:

  • Utilizing improved RNA-Mango variants for enhanced fluorescence intensity and folding stability.
  • Developing a protocol for imaging Mango II tagged RNAs.
  • Applying the protocol to both fixed and live cell imaging.

Main Results:

  • Achieved single-molecule sensitivity in RNA imaging.
  • Demonstrated improved fluorescence intensity and folding stability of RNA-Mango variants.
  • Successfully imaged Mango II tagged RNAs in cellular environments.

Conclusions:

  • Novel RNA-Mango variants significantly advance cellular RNA imaging capabilities.
  • The presented protocol facilitates sensitive and background-free RNA detection.
  • This method opens new avenues for studying RNA dynamics in cells.