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Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling

Our understanding of the mechanisms governing the response to DNA damage in higher eucaryotes crucially depends on our ability to dissect the temporal and spatial organization of the cellular machinery responsible for maintaining genomic integrity. To achieve this goal, we need experimental tools to...

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Autores principales: Ferrando-May, Elisa, Tomas, Martin, Blumhardt, Philipp, Stöckl, Martin, Fuchs, Matthias, Leitenstorfer, Alfred
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712194/
https://www.ncbi.nlm.nih.gov/pubmed/23882280
http://dx.doi.org/10.3389/fgene.2013.00135
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author Ferrando-May, Elisa
Tomas, Martin
Blumhardt, Philipp
Stöckl, Martin
Fuchs, Matthias
Leitenstorfer, Alfred
author_facet Ferrando-May, Elisa
Tomas, Martin
Blumhardt, Philipp
Stöckl, Martin
Fuchs, Matthias
Leitenstorfer, Alfred
author_sort Ferrando-May, Elisa
collection PubMed
description Our understanding of the mechanisms governing the response to DNA damage in higher eucaryotes crucially depends on our ability to dissect the temporal and spatial organization of the cellular machinery responsible for maintaining genomic integrity. To achieve this goal, we need experimental tools to inflict DNA lesions with high spatial precision at pre-defined locations, and to visualize the ensuing reactions with adequate temporal resolution. Near-infrared femtosecond laser pulses focused through high-aperture objective lenses of advanced scanning microscopes offer the advantage of inducing DNA damage in a 3D-confined volume of subnuclear dimensions. This high spatial resolution results from the highly non-linear nature of the excitation process. Here we review recent progress based on the increasing availability of widely tunable and user-friendly technology of ultrafast lasers in the near infrared. We present a critical evaluation of this approach for DNA microdamage as compared to the currently prevalent use of UV or VIS laser irradiation, the latter in combination with photosensitizers. Current and future applications in the field of DNA repair and DNA-damage dependent chromatin dynamics are outlined. Finally, we discuss the requirement for proper simulation and quantitative modeling. We focus in particular on approaches to measure the effect of DNA damage on the mobility of nuclear proteins and consider the pros and cons of frequently used analysis models for FRAP and photoactivation and their applicability to non-linear photoperturbation experiments.
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spelling pubmed-37121942013-07-23 Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling Ferrando-May, Elisa Tomas, Martin Blumhardt, Philipp Stöckl, Martin Fuchs, Matthias Leitenstorfer, Alfred Front Genet Oncology Our understanding of the mechanisms governing the response to DNA damage in higher eucaryotes crucially depends on our ability to dissect the temporal and spatial organization of the cellular machinery responsible for maintaining genomic integrity. To achieve this goal, we need experimental tools to inflict DNA lesions with high spatial precision at pre-defined locations, and to visualize the ensuing reactions with adequate temporal resolution. Near-infrared femtosecond laser pulses focused through high-aperture objective lenses of advanced scanning microscopes offer the advantage of inducing DNA damage in a 3D-confined volume of subnuclear dimensions. This high spatial resolution results from the highly non-linear nature of the excitation process. Here we review recent progress based on the increasing availability of widely tunable and user-friendly technology of ultrafast lasers in the near infrared. We present a critical evaluation of this approach for DNA microdamage as compared to the currently prevalent use of UV or VIS laser irradiation, the latter in combination with photosensitizers. Current and future applications in the field of DNA repair and DNA-damage dependent chromatin dynamics are outlined. Finally, we discuss the requirement for proper simulation and quantitative modeling. We focus in particular on approaches to measure the effect of DNA damage on the mobility of nuclear proteins and consider the pros and cons of frequently used analysis models for FRAP and photoactivation and their applicability to non-linear photoperturbation experiments. Frontiers Media S.A. 2013-07-16 /pmc/articles/PMC3712194/ /pubmed/23882280 http://dx.doi.org/10.3389/fgene.2013.00135 Text en Copyright © 2013 Ferrando-May, Tomas, Blumhardt, Stöckl, Fuchs and Leitenstorfer. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.
spellingShingle Oncology
Ferrando-May, Elisa
Tomas, Martin
Blumhardt, Philipp
Stöckl, Martin
Fuchs, Matthias
Leitenstorfer, Alfred
Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title_full Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title_fullStr Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title_full_unstemmed Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title_short Highlighting the DNA damage response with ultrashort laser pulses in the near infrared and kinetic modeling
title_sort highlighting the dna damage response with ultrashort laser pulses in the near infrared and kinetic modeling
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712194/
https://www.ncbi.nlm.nih.gov/pubmed/23882280
http://dx.doi.org/10.3389/fgene.2013.00135
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