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Related Experiment Video

Updated: Oct 9, 2025

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
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Multilayer Ratiometric Fluorescent Nanomachines for Imaging mRNA in Live Cells.

Jianqiao Chang1,2,3, Yu Zhang4, Yike Li2,3

  • 1Department of Chemistry, Capital Normal University, Beijing, 100048, China.

Small Methods
|December 20, 2021
PubMed
Summary

Researchers developed a novel nanomachine for real-time mRNA detection in live cells. This technology enables precise monitoring of gene silencing during cancer treatment, offering new therapeutic strategies.

Keywords:
live-cell imagingmolecular beaconsnanomachinesiRNA

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Last Updated: Oct 9, 2025

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Area of Science:

  • Biomedical Engineering
  • Molecular Biology
  • Nanotechnology

Background:

  • Detecting mRNA expression in live cells during treatment is crucial for understanding tumor biology and identifying therapeutic targets.
  • Current methods face challenges in real-time monitoring and single-cell resolution.

Purpose of the Study:

  • To develop a multilayer ratiometric fluorescent nanomachine for live-cell perturbation and imaging of mRNA.
  • To enable sequential release of siRNA and molecular beacons for gene silencing and mRNA detection.

Main Methods:

  • Fabrication of nanomachines using microfluidic approaches, featuring fluorescent polymeric cores and multiple lipid layers.
  • Sequential release of siRNA to silence the multidrug resistance 1 (MDR1) gene and molecular beacons to detect MDR1 mRNA.
  • Ratiometric fluorescence measurement for quantifying mRNA levels in MCF-7/ADR cells during treatment.

Main Results:

  • The nanomachines successfully delivered siRNA and molecular beacons to the cytosol in a sequential manner.
  • The fluorescent ratio correlated positively with MDR1 mRNA expression levels during siRNA treatment.
  • The nanomachine's quantification of mRNA showed comparable results to traditional quantitative polymerase chain reaction (qPCR).

Conclusions:

  • The developed nanomachine offers a powerful tool for live-cell perturbation and imaging of mRNA at single-cell resolution.
  • This technology holds significant potential for modulating and imaging intratumoral mRNA in vitro and in vivo.
  • It paves the way for advanced cancer research and the development of novel therapeutic strategies.