Examining DNA Double-Strand Break Repair in a Cell Cycle-Dependent Manner

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

DNA double-strand breaks (DSBs) are deleterious DNA lesions that must be properly repaired to maintain genome stability. Agents, generated both exogenously (environmental radiation, dental X-rays, etc.) and endogenously (reactive oxygen species, DNA replication, V(D)J recombination, etc.), induce numerous DSBs every day. To counter these DSBs, there are two major repair pathways in mammalian cells, nonhomologous end joining (NHEJ) and homologous recombination (HR). NHEJ directly mediates the religation of the broken DNA molecule and is active in all phases of the cell cycle. HR directs repair via the use of a homologous DNA sequence as a template and is primarily active in only S/G2 phases owing to the availability of a DNA template via a sister chromatid. As NHEJ and HR are active in multiple cell cycle phases, there is significant interest in how a cell chooses between the two DSB repair pathways. Therefore, it is essential to utilize assays to study DSB repair that can distinguish between the two DSB repair pathways and the different phases of the cell cycle. In this chapter, we describe methods to measure the contribution of DNA repair pathways in different phases of the cell cycle. These methods are simple, can be applied to most mammalian cell lines, and can be used as a broad utility to monitor cell cycle-dependent DSB repair.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages97-118
Number of pages22
Volume591
DOIs
StatePublished - 2017

Publication series

NameMethods in Enzymology
Volume591
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Cell cycle
  • DNA double-strand breaks
  • DNA end resection
  • EdU labeling
  • Focus formation
  • Homologous recombination
  • Nonhomologous end joining

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Saha, J., Wang, S. Y., & Davis, A. J. (2017). Examining DNA Double-Strand Break Repair in a Cell Cycle-Dependent Manner. In Methods in Enzymology (Vol. 591, pp. 97-118). (Methods in Enzymology; Vol. 591). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2017.03.012