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The study of protein recruitment to laser-induced DNA lesions can be distorted by photoconversion of the DNA binding dye Hoechst.

  • Hurst V Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, Basel, CH-4058, Switzerland.
  • Gasser SM Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, Basel, CH-4058, Switzerland.
  • 2019-04-19
Published in:
  • F1000Research. - 2019
DAPI
DNA repair
Hoechst
Photoconversion
UV laser
Animals
DNA
DNA Repair
Fluorescent Dyes
Lasers
Light
Proteins
English A commonly used approach for assessing DNA repair factor recruitment in mammalian cells is to induce DNA damage with a laser in the UV or near UV range and follow the local increase of GFP-tagged proteins at the site of damage. Often these measurements are performed in the presence of the blue DNA dye Hoechst, which is used as a photosensitizer. However, a light-induced switch of Hoechst from a blue-light to a green-light emitter will give a false positive signal at the site of damage.  Thus, photoconversion signals must be subtracted from the overall green-light emission to determine true recruitment. Here we demonstrate the photoconversion effect and suggest control experiments to exclude false-positive results.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/202687
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