In vivo DNA double-strand breaks enhance gene targeting in cultured silkworm cells

Hiroaki Mon, Takahiro Kusakabe, Jae Man Lee, Yutaka Kawaguchi, Katsumi Koga

研究成果: ジャーナルへの寄稿記事

10 引用 (Scopus)

抄録

Alteration of genomic information through homologous recombination (HR) is a powerful tool for reverse genetics in bacteria, yeast, and mice. The low frequency of HR is, however, a major obstacle to achieve efficient gene targeting. In this study, we have developed an assay system for investigating the frequency of gene targeting in cultured silkworm cells using a firefly luciferase gene as a reporter. The introduction of a DNA double-strand break (DSB) either in the chromosomal target locus or in the targeting construct drastically increased the frequency of gene targeting. Interestingly, the inhibition of poly(ADP-ribose) polymerase (PARP), a protein known to play an important role in overall suppression of the HR pathway, stimulated the targeting efficiency, whereas the overexpression of two silkworm RecA homologs, BmRad51 and BmDmc1, had no effect. The presently devised assay system may serve as a useful tool to improve the gene targeting efficiency in the silkworm (Bombyx mori).

元の言語英語
ページ(範囲)99-106
ページ数8
ジャーナルComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
139
発行部数1
DOI
出版物ステータス出版済み - 9 1 2004

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Bombyx
Gene Targeting
Double-Stranded DNA Breaks
Cultured Cells
Homologous Recombination
Genes
DNA
Assays
Firefly Luciferases
Reverse Genetics
Poly(ADP-ribose) Polymerases
Yeasts
Yeast
Bacteria
Proteins

All Science Journal Classification (ASJC) codes

  • Physiology
  • Biochemistry
  • Molecular Biology

これを引用

In vivo DNA double-strand breaks enhance gene targeting in cultured silkworm cells. / Mon, Hiroaki; Kusakabe, Takahiro; Lee, Jae Man; Kawaguchi, Yutaka; Koga, Katsumi.

:: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 巻 139, 番号 1, 01.09.2004, p. 99-106.

研究成果: ジャーナルへの寄稿記事

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