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

Hiroyuki Kitao, Seiki Hirano, Minoru Takata

研究成果: Contribution to journalArticle査読

3 被引用数 (Scopus)

抄録

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.

本文言語英語
ページ(範囲)293-309
ページ数17
ジャーナルMethods in molecular biology (Clifton, N.J.)
745
DOI
出版ステータス出版済み - 10 4 2011

All Science Journal Classification (ASJC) codes

  • 分子生物学
  • 遺伝学

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