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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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New aromatic cation salts enable photo-induced protein labeling using longer wavelengths of light. These probes offer robust reactivity in complex biological environments and enable wash-free live-cell imaging.

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Area of Science:

  • Chemical Biology
  • Biochemistry
  • Organic Chemistry

Background:

  • Photo-induced protein labeling is crucial in chemical biology.
  • Existing methods often require high-energy light sources.
  • There is a need for probes that utilize longer wavelengths for improved biocompatibility.

Purpose of the Study:

  • Design and synthesize novel aromatic cation salts for photo-induced protein labeling.
  • Investigate structure-reactivity relationships to optimize probe performance.
  • Develop probes capable of functioning with longer wavelengths of light.

Main Methods:

  • Systematic study of donor-acceptor pyridinium salts with extended conjugation.
  • Analysis of structure-reactivity relationships, focusing on the trans-stilbene configuration.
  • Evaluation of probes in complex biological systems, including live-cell imaging and proteomic analysis.

Main Results:

  • Developed aromatic cation salts that trigger protein labeling with longer wavelengths of light.
  • Identified chromene-based donor groups yielding robust labeling and high fluorescence quantum yields.
  • Demonstrated successful green light-triggered intracellular labeling in live HeLa cells, identifying 659 enriched proteins.
  • Probes exhibited state-dependent photophysical properties enabling dual use for labeling and imaging.

Conclusions:

  • This new generation of probes facilitates photo-induced labeling with lower energy light in complex proteomes.
  • The probes offer new capabilities for photophysical state-dependent reactivity and measurements.
  • The developed probes are suitable for both photo-induced protein labeling and wash-free live-cell imaging in biological applications.