In Time and Space: Laser Microirradiation and the DNA Damage Response
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...
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Published in | Methods in molecular biology (Clifton, N.J.) Vol. 1999; p. 61 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
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United States
2019
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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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AbstractList | 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. |
Author | Kumbhar, Ramhari Kim, Jae Jin Miller, Kyle M Gong, Fade |
Author_xml | – sequence: 1 givenname: Jae Jin surname: Kim fullname: Kim, Jae Jin organization: Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA – sequence: 2 givenname: Ramhari surname: Kumbhar fullname: Kumbhar, Ramhari organization: Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA – sequence: 3 givenname: Fade surname: Gong fullname: Gong, Fade organization: Department of Biochemistry & Molecular Biology, Baylor College of Medicine, Houston, TX, USA – sequence: 4 givenname: Kyle M surname: Miller fullname: Miller, Kyle M email: kyle.miller@austin.utexas.edu organization: Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA. kyle.miller@austin.utexas.edu |
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CitedBy_id | crossref_primary_10_1016_j_molcel_2020_08_018 crossref_primary_10_1016_j_dnarep_2023_103592 crossref_primary_10_1093_nar_gkac160 crossref_primary_10_3389_fcell_2022_920267 crossref_primary_10_1016_j_mrrev_2021_108397 crossref_primary_10_1083_jcb_202006149 |
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Keywords | Laser microirradiation Live-cell imaging Transcription DNA repair DNA damage Fluorescence microscopy |
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Snippet | Maintenance of genomic integrity depends on the spatiotemporal recruitment and regulation of DNA damage response and repair proteins at DNA damage sites. These... |
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SubjectTerms | Cell Line, Tumor DNA Damage - radiation effects DNA Repair - radiation effects DNA-Binding Proteins - chemistry DNA-Binding Proteins - metabolism Epigenome - radiation effects Fluorescent Dyes - chemistry Genomic Instability - radiation effects Humans Intravital Microscopy - instrumentation Intravital Microscopy - methods Lasers Microscopy, Confocal - instrumentation Microscopy, Confocal - methods Microscopy, Fluorescence - instrumentation Microscopy, Fluorescence - methods Spatio-Temporal Analysis Transcription, Genetic - radiation effects |
Title | In Time and Space: Laser Microirradiation and the DNA Damage Response |
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