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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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Conditionally Activated ROS Generation by MMP2/9-Specific C60-Based Fluorescence Probes.

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  • 1Department of Chemistry and Applied Biosciences, ETH Zürich, Vladimir-Prelog-Weg 3, Zürich, 8093, Switzerland.

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A novel C60-coumarin probe detects matrix metalloproteinase-2/9 (MMP2/9) in cancer cells. Upon MMP2/9 cleavage, the probe enhances fluorescence and photodynamic therapy for targeted cancer treatment.

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Area of Science:

  • Bioconjugate Chemistry
  • Molecular Imaging
  • Photodynamic Therapy

Background:

  • Matrix metalloproteinases (MMPs), particularly MMP2 and MMP9, are implicated in tumor progression and metastasis.
  • Developing targeted probes for MMPs is crucial for cancer diagnosis and therapy.
  • Existing probes often lack sufficient specificity or dual functionality for imaging and treatment.

Purpose of the Study:

  • To design and synthesize a C60-coumarin conjugate probe for detecting MMP2/9 activity.
  • To evaluate the probe's capability for fluorescence imaging of MMP2/9-expressing cells.
  • To assess the probe's potential as a photosensitizer for photodynamic therapy in cancer.

Main Methods:

  • Synthesis of a C60-coumarin conjugate linked by an MMP2/9-cleavable peptide.
  • Characterization of fluorescence quenching and recovery upon peptide cleavage.
  • In vitro cellular imaging experiments using tumor cell lines with varying MMP2/9 expression levels.
  • Assessment of photodynamic therapy efficacy based on MMP2/9 activity.

Main Results:

  • The C60-coumarin probe exhibited quenched fluorescence, which was restored upon cleavage by MMP2/9.
  • Specificity was confirmed using a scrambled peptide control and an MMP2 inhibitor.
  • Successful in vitro imaging of MMP2/9 in high-expression DAOY cells was achieved.
  • Enhanced 1O2 generation and photocytotoxicity were observed in MMP2/9-expressing cells.

Conclusions:

  • The developed probe functions as a specific fluorescent reporter for MMP2/9.
  • The probe acts as an effective photosensitizer for MMP2/9-dependent photodynamic therapy.
  • This dual-function probe holds promise for targeted cancer imaging and treatment.