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DNA Repair pp 61–74Cite as

In Time and Space: Laser Microirradiation and the DNA Damage Response

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1999))

Abstract

Maintenance of genomic integrity depends on the spatiotemporal recruitment and regulation of DNA damage response and repair proteins at DNA damage sites. These highly dynamic processes have been widely studied using laser microirradiation coupled with fluorescence microscopy. Laser microirradiation has provided a powerful methodology to identify and determine mechanisms of DNA damage response pathways. Here we describe methods used to analyze protein recruitment dynamics of fluorescently tagged or endogenous proteins to laser-induced DNA damage sites using laser scanning and fluorescence microscopy. We further describe multiple applications employing these techniques to study additional processes at DNA damage sites including transcription. Together, we aim to provide robust visualization methods employing laser-microirradiation to detect and determine protein behavior, functions and dynamics in response to DNA damage in mammalian cells.

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Acknowledgments

The K.M.M. laboratory is supported by the NIH National Cancer Institute (R01CA198279 and RO1CA201268) and the American Cancer Society (RSG-16-042-01-DMC).

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Authors and Affiliations

  1. Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA

    Jae Jin Kim, Ramhari Kumbhar, Fade Gong & Kyle M. Miller

  2. Department of Biochemistry & Molecular Biology, Baylor College of Medicine, Houston, TX, USA

    Fade Gong

Authors
  1. Jae Jin Kim
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  2. Ramhari Kumbhar
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  3. Fade Gong
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  4. Kyle M. Miller
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Corresponding author

Correspondence to Kyle M. Miller .

Editor information

Editors and Affiliations

  1. Indiana University – Purdue University, Indianapolis, IN, USA

    Lata Balakrishnan

  2. Department of Biological Sciences, University of South Carolina, Columbia, SC, USA

    Jason A. Stewart

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Kim, J.J., Kumbhar, R., Gong, F., Miller, K.M. (2019). In Time and Space: Laser Microirradiation and the DNA Damage Response. In: Balakrishnan, L., Stewart, J. (eds) DNA Repair. Methods in Molecular Biology, vol 1999. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9500-4_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9500-4_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9499-1

  • Online ISBN: 978-1-4939-9500-4

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