Evaluation of homologous recombinational repair in chicken B lymphoma cell line, DT40.

Hiroyuki Kitao, Seiki Hirano, Minoru Takata

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)293-309
Number of pages17
JournalMethods in molecular biology (Clifton, N.J.)
Volume745
DOIs
Publication statusPublished - Oct 4 2011

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Recombinational DNA Repair
Chickens
Lymphoma
Cell Line
Homologous Recombination
Vertebrates
Hereditary Neoplastic Syndromes
Genes
Fanconi Anemia
Sister Chromatid Exchange
Gene Targeting
Deoxyribonuclease I
Gene Frequency
Cell Survival
Yeasts
Phenotype
Mutation
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Evaluation of homologous recombinational repair in chicken B lymphoma cell line, DT40. / Kitao, Hiroyuki; Hirano, Seiki; Takata, Minoru.

In: Methods in molecular biology (Clifton, N.J.), Vol. 745, 04.10.2011, p. 293-309.

Research output: Contribution to journalArticle

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