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γH2AX foci analysis for monitoring DNA double-strand break repair: Strengths, limitations and optimization

Löbrich, Markus ; Shibata, Atsushi ; Beucher, Andrea ; Fisher, Anna ; Ensminger, Michael ; Goodarzi, Aaron A. ; Barton, Olivia ; Jeggo, Penny A. (2021)
γH2AX foci analysis for monitoring DNA double-strand break repair: Strengths, limitations and optimization.
In: Cell Cycle, 2010, 9 (4)
doi: 10.26083/tuprints-00019060
Artikel, Zweitveröffentlichung, Verlagsversion

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Kurzbeschreibung (Abstract)

DNA double-strand breaks (DSBs) represent an important radiation-induced lesion and impaired DSB repair provides the best available correlation with radiosensitivity. Physical techniques for monitoring DSB repair require high, non-physiological doses and cannot reliably detect subtle defects. One outcome from extensive research into the DNA damage response is the observation that H2AX, a variant form of the histone H2A, undergoes extensive phosphorylation at the DSB, creating γH2AX foci that can be visualised by immunofluorescence. There is a close correlation between γH2AX foci and DSB numbers and between the rate of foci loss and DSB repair, providing a sensitive assay to monitor DSB repair in individual cells using physiological doses. However, γH2AX formation can occur at single-stranded DNA regions which arise during replication or repair and thus does not solely correlate with DSB formation. Here, we present and discuss evidence that following exposure to ionising radiation, γH2AX foci analysis can provide a sensitive monitor of DSB formation and repair and describe techniques to optimise the analysis. We discuss the limitations and benefits of the technique, enabling the procedure to be optimally exploited but not misused.

Typ des Eintrags: Artikel
Erschienen: 2021
Autor(en): Löbrich, Markus ; Shibata, Atsushi ; Beucher, Andrea ; Fisher, Anna ; Ensminger, Michael ; Goodarzi, Aaron A. ; Barton, Olivia ; Jeggo, Penny A.
Art des Eintrags: Zweitveröffentlichung
Titel: γH2AX foci analysis for monitoring DNA double-strand break repair: Strengths, limitations and optimization
Sprache: Englisch
Publikationsjahr: 2021
Publikationsdatum der Erstveröffentlichung: 2010
Verlag: Taylor and Francis Group
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Cell Cycle
Jahrgang/Volume einer Zeitschrift: 9
(Heft-)Nummer: 4
DOI: 10.26083/tuprints-00019060
URL / URN: https://tuprints.ulb.tu-darmstadt.de/19060
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Herkunft: Zweitveröffentlichungsservice
Kurzbeschreibung (Abstract):

DNA double-strand breaks (DSBs) represent an important radiation-induced lesion and impaired DSB repair provides the best available correlation with radiosensitivity. Physical techniques for monitoring DSB repair require high, non-physiological doses and cannot reliably detect subtle defects. One outcome from extensive research into the DNA damage response is the observation that H2AX, a variant form of the histone H2A, undergoes extensive phosphorylation at the DSB, creating γH2AX foci that can be visualised by immunofluorescence. There is a close correlation between γH2AX foci and DSB numbers and between the rate of foci loss and DSB repair, providing a sensitive assay to monitor DSB repair in individual cells using physiological doses. However, γH2AX formation can occur at single-stranded DNA regions which arise during replication or repair and thus does not solely correlate with DSB formation. Here, we present and discuss evidence that following exposure to ionising radiation, γH2AX foci analysis can provide a sensitive monitor of DSB formation and repair and describe techniques to optimise the analysis. We discuss the limitations and benefits of the technique, enabling the procedure to be optimally exploited but not misused.

Status: Verlagsversion
URN: urn:nbn:de:tuda-tuprints-190607
Sachgruppe der Dewey Dezimalklassifikatin (DDC): 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Radiation Biology and DNA Repair
Hinterlegungsdatum: 10 Sep 2021 12:06
Letzte Änderung: 13 Sep 2021 06:14
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