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Homologous Recombination Assay for Interstrand Cross-Link Repair

  • Koji Nakanishi
  • Francesca Cavallo
  • Erika Brunet
  • Maria JasinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 745)

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.

Key words

Homologous recombination interstrand cross-link repair triplex-forming oligonucleotide GFP reporters 

Notes

Acknowledgments

This work was supported by the Byrne Fund and National Institutes for Health grants P01CA94060 (M.J.) and R01GM54668 (M.J.).

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Koji Nakanishi
    • 1
  • Francesca Cavallo
    • 2
  • Erika Brunet
    • 3
  • Maria Jasin
    • 1
    Email author
  1. 1.Developmental Biology ProgramMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  2. 2.Department of Public Health and Cell Biology, Section of AnatomyUniversity of Rome Tor VergataRomeItaly
  3. 3.Muséum National d’Histoire NaturelleParisFrance

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