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

Markus Löbrich, Atsushi Shibata, Andrea Beucher, Anna Fisher, Michael Ensminger, Aaron A. Goodarzi, Olivia Barton, Penny A. Jeggo

Research output: Contribution to journalReview articlepeer-review

525 Citations (Scopus)

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 visualized by immuno-fluorescence. 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 ionizing radiation, γH2AX foci analysis can provide a sensitive monitor of DSB formation and repair and describe techniques to optimize the analysis. We discuss the limitations and benefits of the technique, enabling the procedure to be optimally exploited but not misused.

Original languageEnglish
Pages (from-to)662-669
Number of pages8
JournalCell Cycle
Volume9
Issue number4
DOIs
Publication statusPublished - 2010 Feb 15
Externally publishedYes

Keywords

  • Ataxia telangiectasia
  • Double-strand break repair
  • Hydrogen peroxide
  • Ionizing radiation
  • Pulsed-field gel electrophoresis
  • Single-stranded DNA
  • γH2AX foci

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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