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DNA Double-Strand Break Repair Assay

Chen-Chun Pai1, Elizabeth Blaikley1, Timothy C Humphrey1

  • 1CRUK-MRC Institute for Radiation Oncology, University of Oxford, Department of Oncology, ORCRB, Oxford OX3 7DQ, United Kingdom.

Cold Spring Harbor Protocols
|July 23, 2017
PubMed

Insights

This study presents a new DNA double-strand break (DSB) assay in fission yeast. The assay quantifies DNA repair, misrepair, and failed repair events, aiding genome stability research.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA double-strand breaks (DSBs) are critical DNA lesions that can cause genome instability and cell death if unrepaired.
  • Understanding the mechanisms of DSB repair and misrepair is crucial for both fundamental biology and medicine.

Purpose of the Study:

  • To describe a novel assay for identifying and quantifying DNA double-strand break (DSB) repair, misrepair, and failed repair events.
  • To enable the study of factors influencing DSB repair and genome stability in *Schizosaccharomyces pombe*.

Main Methods:

  • Development of a site-specific DSB assay using a nonessential minichromosome (Ch16) in *Schizosaccharomyces pombe*.
  • Quantification of various outcomes of DSB repair, including accurate repair, misrepair, and failed repair.

Main Results:

  • The assay successfully identifies and quantifies different DSB repair and misrepair events.
  • This method allows for the assessment of gene contributions to DSB repair and genome stability.

Conclusions:

  • The developed DSB assay is a valuable tool for investigating DNA repair pathways and genome stability.
  • The assay provides mechanistic insights into the function of genes involved in DSB repair.

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