DNA Repair pp 61-74 | Cite as

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

  • Jae Jin Kim
  • Ramhari Kumbhar
  • Fade Gong
  • Kyle M. MillerEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1999)


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.

Key words

Laser microirradiation DNA repair DNA damage Transcription Fluorescence microscopy Live-cell imaging 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jae Jin Kim
    • 1
  • Ramhari Kumbhar
    • 1
  • Fade Gong
    • 1
    • 2
  • Kyle M. Miller
    • 1
    Email author
  1. 1.Department of Molecular Biosciences, Institute for Cellular and Molecular BiologyThe University of Texas at AustinAustinUSA
  2. 2.Department of Biochemistry & Molecular BiologyBaylor College of MedicineHoustonUSA

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