Homologous recombination assay for interstrand cross-link repair

Methods Mol Biol. 2011;745:283-91. doi: 10.1007/978-1-61779-129-1_16.

Abstract

DNA interstrand cross-links (ICLs) covalently link both strands of the DNA duplex, impeding cellular processes like DNA replication. Homologous recombination (HR) is considered to be a major pathway for the repair of ICLs in mammalian cells as mutants for HR components are highly sensitive to DNA-damaging agents that cause ICLs. This chapter describes GFP assays to measure HR following site-specific ICL formation with psoralen through DNA triplex technology. This approach can be used to determine the genetic requirements for ICL-induced HR in relation to those involved in HR repair of other DNA lesions such as double-strand breaks.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line, Tumor
  • DNA Repair / genetics*
  • Flow Cytometry
  • Humans
  • Oligonucleotides / genetics
  • Plasmids / genetics
  • Recombination, Genetic / genetics*

Substances

  • Oligonucleotides