Multimodal Detection and Quantification of DNA Double Strand Breaks (DSBs) in Cells Using Plate Readers
Prof. Felicite Noubissi-Kamdem
Department of Biology/RCMI-CHDR
Jackson State University
Radiotherapy uses γ irradiation to induce DNA double strand breaks (DSBs) leading to cell death if not repaired. These DNA double strand breaks are always followed by phosphorylation of the histone protein component H2AX to form γ-H2AX. γ-H2AX was shown to represent DSBs in a 1:1 manner and has been therefore established as a marker for DNA DSBs. Accurate quantifying γ-H2AX as an indicator of DNA DSBs could help improve the proper dosage of precision radiation and chemotherapy. We have developed a multimodal detection of γ-H2AX foci in a 2D cell culture system to quantify DNA DSBs in cells using a plate reader. This multimodal detection approach measures the fluorescence intensity of γ-H2AX labeled with the traditional fluorophore Alexa Fluor 488, or europium- labeled secondary antibody in the dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) that employs time resolved fluorescence (TRF) intensity technology. These approaches have the potential to improve precision chemo and radiotherapy, assess the protective effect of some compounds against ionizing radiation, or quantify DNA damage induced by chemicals and physical factors.
Hosted by Dr. deBoer