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Analyzing Heterochromatic DNA Double Strand Break (DSB) Repair in Response to Ionizing Radiation

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ATM Kinase

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1599))

Abstract

DNA damaging agents such as ionizing irradiation induce lesions in the DNA such as double strand breaks (DSBs). Depending on cell type, 10–25% of these DSBs are induced in heterochromatin. Heterochromatic DSBs are resolved with slow kinetics (compared to DSBs in euchromatin) and require ATM activity for repair. Investigating the underlying causes of the slow component of DSB repair and the role of individual response factors in this process provides insight into DSB response pathways and will further the understanding of diseases where such pathways are dysfunctional due to mutation. Here, we describe a method to detect DSB repair foci in the heterochromatin of human cells. We provide a detailed protocol for cell culture preparation, immunofluorescence microscopy, and a computer-assisted approach to analyze overlap between DSB foci and heterochromatin.

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Authors and Affiliations

  1. Robson DNA Science Centre, Arnie Charbonneau Cancer Institute, Department of Biochemistry and Molecular Biology and Department of Oncology, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada, T2N 4N1

    Karolin Klement & Aaron A. Goodarzi

Authors
  1. Karolin Klement
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  2. Aaron A. Goodarzi
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Correspondence to Aaron A. Goodarzi .

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Editors and Affiliations

  1. University of Queensland Centre for Clinical Research (UQCCR), University of Queensland, Herston, Queensland, Australia

    Sergei V. Kozlov

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Klement, K., Goodarzi, A.A. (2017). Analyzing Heterochromatic DNA Double Strand Break (DSB) Repair in Response to Ionizing Radiation. In: Kozlov, S. (eds) ATM Kinase. Methods in Molecular Biology, vol 1599. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6955-5_22

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  • DOI: https://doi.org/10.1007/978-1-4939-6955-5_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6953-1

  • Online ISBN: 978-1-4939-6955-5

  • eBook Packages: Springer Protocols

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