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Related Concept Videos

Reporter Genes02:11

Reporter Genes

11.3K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Labeling DNA Probes03:31

Labeling DNA Probes

8.2K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Related Experiment Video

Updated: Jun 30, 2025

A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence FUEL
07:04

A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence FUEL

Published on: May 23, 2014

11.6K

FLEX: genetically encodable enzymatic fluorescence signal amplification using engineered peroxidase.

Nirmali Sharma1, Minkyo Jung2, Pratyush Kumar Mishra3

  • 1Department of Chemistry, Seoul National University, Seoul 08826, Korea; Department of Chemistry, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Korea.

Cell Chemical Biology
|March 21, 2024
PubMed
Summary

We developed FLEX, a new method using enzymatic fluorescence signal amplification, to visualize biomolecules within cells. This technique enhances fluorescence detection and preserves cellular structures for advanced biological imaging.

Keywords:
APEXCLEMFLEXcorrelative light and electron microscopyfluorescent peroxidase substratefluorescent probefluorescent probeslysosomemitochondriaorganelle communicationproximity labeling

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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

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Last Updated: Jun 30, 2025

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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Microscopy

Background:

  • Peroxidase-based reactions are used for signal amplification in biomolecule detection.
  • Genetically encodable enzymatic methods offer potential for sensitive cellular analysis.

Purpose of the Study:

  • To develop a genetically encodable enzymatic fluorescence signal amplification method (FLEX) for sensitive detection of biomolecules.
  • To enable fluorescence-based detection of subcellularly localized metabolites and proteins of interest (POIs).

Main Methods:

  • Developed FLEX, a method utilizing the APEX enzyme and a novel fluorescent probe (Jenfluor triazole, JFT1).
  • Synthesized JFT1 for amplified and restricted fluorescence signals under fixed conditions.
  • Applied FLEX to various APEX-conjugated POIs targeted to different organelles.

Main Results:

  • FLEX enabled sensitive, fluorescence-based detection of subcellularly localized metabolites.
  • JFT1 demonstrated stable fluorescence signals under osmium treatment and polymer embedding, supporting correlative light and electron microscopy (CLEM).
  • FLEX successfully visualized POI localization while preserving organelle ultrastructures and provided insights into lysosome-mitochondria interactions.

Conclusions:

  • FLEX is a sensitive and versatile system for fluorescently detecting APEX2-POIs in multiscale biological samples.
  • The method allows for effective visualization of cellular components and dynamic interactions.
  • FLEX holds significant promise for advancing biological imaging and analysis.