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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

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

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Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics
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Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics

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DNA-Barcoded Plasmonic Nanostructures for Activity-Based Protease Sensing.

Subrata Pandit1, Mark Duchow2, Wilson Chao3

  • 1Department of Chemistry, The University of Texas at Austin, 105 E 24th St., Austin, TX 78712, USA.

Angewandte Chemie (International Ed. in English)
|November 20, 2023
PubMed
Summary
This summary is machine-generated.

We developed novel DNA-barcoded plasmonic nanostructures for sensitive protease detection. These sensors offer high specificity and multiplexing capabilities for disease diagnostics.

Keywords:
Activity-Based SensingDNAPeptidePlasmonic NanostructureProtease

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Area of Science:

  • Nanotechnology
  • Biochemistry
  • Molecular Biology

Background:

  • Protease activity is crucial in various biological processes and disease states.
  • Existing sensors often lack sensitivity, specificity, or multiplexing capabilities for complex diagnostics.

Purpose of the Study:

  • To develop a new class of highly sensitive and multiplexed protease activity sensors.
  • To demonstrate the utility of DNA-barcoded plasmonic nanostructures (GPDs) for detecting clinically relevant proteases.

Main Methods:

  • Functionalization of gold nanoparticles with peptide-DNA conjugates (GPDs) as protease substrates.
  • CRISPR/Cas12a-based assay for detecting protease-cleaved DNA barcodes.
  • Colorimetric detection via nanoparticle aggregation for visual readout.

Main Results:

  • GPDs achieved >25-fold signal turn-on and 100-fold improved response over commercial probes.
  • Detection limits as low as 58 pM at room temperature with high sensitivity and selectivity.
  • Successful multiplexed detection of SARS-CoV-2 3CL protease and caspase 3, and visual detection of cathepsin B in patient-derived cell lines.

Conclusions:

  • DNA-barcoded plasmonic nanostructures (GPDs) provide a powerful platform for sensitive and multiplexed protease detection.
  • GPDs offer significant advantages in biosensing and disease diagnostics, including minimal sample processing and visual readout capabilities.
  • This technology holds promise for detecting physiologically relevant protease concentrations in complex biological samples for clinical applications.