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Examining the Role of HDACs in DNA Double-Strand Break Repair in Neurons

  • Ping-Chieh Pao
  • Jay Penney
  • Li-Huei TsaiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1983)

Abstract

Histone deacetylases (HDACs) modulate chromatin structure by removing acetyl groups from histones. Upon DNA double-strand breaks (DSBs), deacetylation of H3K56 and H4K16 by HDACs occurs immediately at break sites, and is crucial for DSB repair. Here we describe two assays that examine defective DSB repair caused by HDAC inhibition in primary cortical neurons: single-cell gel electrophoresis to assay DNA integrity (the comet assay) and western blot analysis for γH2AX, a phosphorylated histone variant associated with DSBs.

Keywords

Genome stability DNA damage Histone deacetylation Histone deacetylases (HDACs) DNA repair DNA double-strand breaks (DSBs) Single-cell gel electrophoresis assay Comet assay γH2AX 

Notes

Acknowledgments

This work was supported by NIA grant (AG046174), NINDS grant (NS102730), and Glenn award for research in biological mechanism of aging.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Picower Institute for Learning and MemoryMassachusetts Institute of TechnologyCambridgeUSA
  2. 2.Department of Brain and Cognitive SciencesMassachusetts Institute of TechnologyCambridgeUSA

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