Mapping DNA Breaks by Next-Generation Sequencing

Methods Mol Biol. 2018:1672:155-166. doi: 10.1007/978-1-4939-7306-4_13.

Abstract

Here, we present two approaches to map DNA double-strand breaks (DSBs) and single-strand breaks (SSBs) in the genome of human cells. We named these methods respectively DSB-Seq and SSB-Seq. We tested the DSB and SSB-Seq in HCT1116, human colon cancer cells, and validated the results using the topoisomerase 2 (Top2)-poisoning agent etoposide (ETO). These methods are powerful tools for the direct detection of the physiological and pathological "breakome" of the DNA in human cells.

Keywords: DNA damage; Double-strand breaks (DSBs); Etoposide (ETO); Single-strand breaks (SSBs); Topoisomerase 2 (Top2).

MeSH terms

  • Chromosome Mapping* / methods
  • Computational Biology / methods
  • DNA / chemistry
  • DNA / genetics
  • DNA / isolation & purification
  • DNA Breaks, Double-Stranded* / drug effects
  • DNA Damage
  • DNA Topoisomerases, Type II / metabolism
  • DNA, Single-Stranded
  • Etoposide / pharmacology
  • High-Throughput Nucleotide Sequencing* / methods
  • Sequence Analysis, DNA

Substances

  • DNA, Single-Stranded
  • Etoposide
  • DNA
  • DNA Topoisomerases, Type II