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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

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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: Dec 26, 2025

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

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A DNA-based fluorescent probe maps NOS3 activity with subcellular spatial resolution.

Maulik S Jani1,2, Junyi Zou1,2, Aneesh T Veetil1,2

  • 1Department of Chemistry, University of Chicago, Chicago, IL, USA.

Nature Chemical Biology
|March 11, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed NOckout probes to map nitric oxide synthase 3 (NOS3) activity in live cells. This technology revealed NOS3

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Mapping Absolute DNA Density in Cell Nuclei using Single-molecule Localization Microscopy
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Probing Structural and Dynamic Properties of Trafficking Subcellular Nanostructures by Spatiotemporal Fluctuation Spectroscopy

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Nitric oxide synthase 3 (NOS3) generates nitric oxide (NO), a crucial signaling molecule involved in protein S-nitrosylation.
  • Endogenous NOS3 is localized to the plasma membrane and the trans-Golgi network (TGN), but its distinct activities and contributions are not fully understood.

Purpose of the Study:

  • To develop a method for spatially mapping the activity of endogenous NOS3 at the plasma membrane and TGN in live cells.
  • To investigate the relative contributions of different NOS3 pools to cellular functions.

Main Methods:

  • Development of a fluorescent DNA-based probe technology, termed NOckout.
  • Targeting NOckout probes to specific subcellular locations (plasma membrane and TGN).
  • Quantitative mapping of endogenous NOS3 activity in live cells using the NOckout system.

Main Results:

  • NOS3 activity at the Golgi is tenfold less active compared to the plasma membrane.
  • Despite lower activity, NOS3 at the Golgi is essential for maintaining Golgi structural integrity.
  • The study demonstrates the capability to spatially resolve NOS3 activity in live cells.

Conclusions:

  • The NOckout probe technology enables quantitative mapping of NOS3 activity at distinct subcellular locations.
  • NOS3 activity within the Golgi plays a critical role in Golgi structure.
  • This platform facilitates the discovery of selective regulators for different NOS3 pools.