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Related Concept Videos

FISH - Fluorescent In-situ Hybridization02:07

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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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Updated: May 21, 2025

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Imaging Low-Abundance, Short Cellular RNAs Using G-quadruplex FISH.

Yong Zhang1, Xinlei Zhang1, Chaoshuang Qu1

  • 1College of Biomass Science and Engineering, Sichuan University, Chengdu, Sichuan 610065, China.

Nano Letters
|May 19, 2025
PubMed
Summary

G-quadruplex FISH enhances RNA imaging sensitivity by using tyramide deposition for signal amplification. This method improves detection of low-abundance and short RNAs, aiding cellular function and disease research.

Keywords:
G-quadruplex peroxidaseendotoxinfluorescence in situ hybridizationshort RNAssingle-cell imaging

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Visualizing cellular RNAs offers spatial resolution at the single-cell level.
  • Conventional fluorescence in situ hybridization (FISH) lacks sensitivity for low-abundance or short RNAs due to limited signal gain.

Purpose of the Study:

  • To develop a sensitive RNA imaging method to overcome the limitations of conventional FISH.
  • To enable the detection and visualization of low-abundance and short RNAs in cells.

Main Methods:

  • Development of G-quadruplex FISH, a method combining G-quadruplex peroxidase-driven tyramide deposition with proximity labeling.
  • Utilizing catalytic tyramide polymerization for signal amplification without nucleic acid amplification.
  • Assessing signal-to-background ratio and RNA detection limits.

Main Results:

  • G-quadruplex FISH significantly reduces background and enhances signal-to-background ratio compared to conventional FISH.
  • Achieved sensitive detection of RNAs down to 25.7 copies/cell.
  • Enabled visualization of short RNAs and exploration of endotoxin immunotoxicity and antagonist screening.

Conclusions:

  • G-quadruplex FISH overcomes the sensitivity limitations of traditional FISH.
  • This technique facilitates the investigation of diverse RNA roles in cellular functions and diseases.
  • Provides a powerful tool for high-gain RNA imaging in biological research.