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Updated: Jun 1, 2026

Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells Using GFP-Based Reporter Systems
Published on: February 2, 2024
Homologous recombination assay for interstrand cross-link repair
Koji Nakanishi1, Francesca Cavallo, Erika Brunet
1Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. nakanisk@mskcc.org
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.
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