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Quantifying kinetics and dynamics of DNA repair proteins using Raster-scan Image Correlation Spectroscopy and Fluorescence Recovery after Photobleaching

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Abstract

DNA double-strand breaks (DSBs) are one of the most lethal DNA damage occurs in mammalian cells. Improper repair of DSBs may leads to mutations, chromosomal translocations, apoptosis, genetic instability [1, 2], increases the chance to cancer development [3] and immune deficiency [4]. In this study the confocal Raster scan Image Correlation Spectroscopy technique is used to study kinetics and dynamics of double stand break repair proteins after γ-irradiation of mammalian cells. Diffusion and binding constants were obtained by fitting with different physical models. Results were compared to ones obtained by creating high density DNA damage with a laser and subsequently performing Fluorescence Recovery after Photobleaching over the damage area. This work presents similarities and differences in double strand break repair response between γ-irradiation versus laser damage.

© 2011 Optical Society of America

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