title: Evaluation of homologous recombinational repair in chicken B lymphoma cell line, DT40.
abstract: Homologous recombination (HR) is a mode of double-strand break (DSB) repair required for cell viability in vertebrate cells. Targeted integration of homologous DNA fragment by HR is usually a very rare event in vertebrate cells; however, in chicken B lymphoma cell line DT40, the ratio of targeted to random integration is extremely high. Although the underlying mechanism of this phenotype is not fully understood, DT40 has been utilized as a model cell line for a number of genetic analyses. Here we describe three assays for evaluating homologous recombinational repair (HRR) using DT40 as a model system, measuring gene-targeting frequency, analyzing HRR process of single DSB induced by yeast homing endonuclease I-SceI, and measuring sister chromatid exchange frequency. Combined with generation of gene-disrupted DT40 mutant cell line, these assays have been highly useful to investigate the mechanisms in HRR. Using these techniques, a role of HRR of not only Rad52 epistasis group genes but also genes whose mutation cause hereditary cancer syndrome, such as Fanconi anemia, has been established.
authors.affiliation: Department of Molecular Oncology, Kyushu University, Kyushu, Japan. hkitao@surg2.med.kyushu-u.ac.jp
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kitao
authors.forename: Seiki
authors.initials: S
authors.lastname: Hirano
authors.forename: Minoru
authors.initials: M
authors.lastname: Takata
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002645:Chickens; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair / Q000235:genetics*; D016393:Lymphoma, B-Cell / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics*; D012854:Sister Chromatid Exchange / Q000235:genetics