PMID- 20139725
OWN - NLM
STAT- MEDLINE
DCOM- 20100608
LR  - 20220409
IS  - 1551-4005 (Electronic)
IS  - 1551-4005 (Linking)
VI  - 9
IP  - 4
DP  - 2010 Feb 15
TI  - gammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, 
      limitations and optimization.
PG  - 662-9
AB  - 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 gammaH2AX foci that can be visualized by immunofluorescence. 
      There is a close correlation between gammaH2AX 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, gammaH2AX 
      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, gammaH2AX 
      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.
FAU - Lobrich, Markus
AU  - Lobrich M
AD  - Darmstadt University of Technology, Radiation Biology and DNA Repair, Darmstadt, 
      Germany. lobrich@bio.tu-darmstadt.de
FAU - Shibata, Atsushi
AU  - Shibata A
FAU - Beucher, Andrea
AU  - Beucher A
FAU - Fisher, Anna
AU  - Fisher A
FAU - Ensminger, Michael
AU  - Ensminger M
FAU - Goodarzi, Aaron A
AU  - Goodarzi AA
FAU - Barton, Olivia
AU  - Barton O
FAU - Jeggo, Penny A
AU  - Jeggo PA
LA  - eng
GR  - G0500897/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
DEP - 20100302
PL  - United States
TA  - Cell Cycle
JT  - Cell cycle (Georgetown, Tex.)
JID - 101137841
RN  - 0 (DNA, Single-Stranded)
RN  - 0 (H2AX protein, human)
RN  - 0 (Histones)
RN  - BBX060AN9V (Hydrogen Peroxide)
SB  - IM
MH  - Cell Line
MH  - *DNA Breaks, Double-Stranded/drug effects/radiation effects
MH  - *DNA Repair
MH  - DNA, Single-Stranded
MH  - G1 Phase
MH  - G2 Phase
MH  - Histones/*analysis/metabolism
MH  - Humans
MH  - Hydrogen Peroxide/pharmacology
MH  - Microscopy, Fluorescence
MH  - Radiation, Ionizing
MH  - S Phase
EDAT- 2010/02/09 06:00
MHDA- 2010/06/09 06:00
CRDT- 2010/02/09 06:00
PHST- 2010/02/09 06:00 [entrez]
PHST- 2010/02/09 06:00 [pubmed]
PHST- 2010/06/09 06:00 [medline]
AID - 10764 [pii]
AID - 10.4161/cc.9.4.10764 [doi]
PST - ppublish
SO  - Cell Cycle. 2010 Feb 15;9(4):662-9. doi: 10.4161/cc.9.4.10764. Epub 2010 Mar 2.