The Gene Cluster Instability (GCI) Assay for Recombination

Methods Mol Biol. 2020:2102:459-482. doi: 10.1007/978-1-0716-0223-2_26.

Abstract

A newly developed method for quantitatively detecting genomic restructuring in cultured human cell lines as the result of recombination is presented: the "gene cluster instability" (GCI) assay. The assay is physiological in that it detects spontaneous restructuring without the need for exogenous recombination-initiating treatments such as DNA damage. As an assay for genotoxicity, the GCI assay is complementary to well-established sister chromatid exchange (SCE) methods. Analysis of the U-2 OS osteosarcoma cell line is presented as an illustration of the method.

Keywords: Gene cluster; Genomic instability; Genotoxicity; Recombination.

MeSH terms

  • Blotting, Southern
  • Cell Line, Tumor
  • DNA / drug effects
  • DNA / genetics
  • DNA / isolation & purification
  • DNA Restriction Enzymes
  • Electrophoresis, Agar Gel
  • Genomic Instability
  • Humans
  • Multigene Family
  • Mutagenicity Tests / methods*
  • Recombination, Genetic*
  • Workflow

Substances

  • DNA
  • DNA Restriction Enzymes