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RecA-mediated, targeted mutagenesis in zebrafish
Zongbin Cui; Ying Yang; Christopher D. Kaufman; Dritan Agalliu; Perry B. Hackett; Yang, Y, Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA
AbstractWe have evaluated the efficacy of RecA, a prokaryotic protein involved with homologous recombination, to direct site-specific mutagenesis in zebrafish embryos. For this we coinjected a vector containing a mutated enhanced green fluorescent protein (EGFP) gene plus 236-nucleotide corrective single-stranded DNAs coated with RecA into I-cell zebrafish embryos. Twenty-hours after fertilization, about 5% to 20% of injected embryos showed EGFP expression in I or more cells when RecA-coated corrective DNAs were used, but not when RecA was omitted. Mutated EGFP genes with 1-bp insertions or deletions were inefficiently activated, whereas those with 7-bp insertions were activated about 4-fold more efficiently. RecA-coated template strand had a higher efficiency than its complementary strand in activation of EGFP expression. Prior irradiation of the embryos with UV light enhanced RecA-mediated restoration of gene activity, suggesting that the effects we observed were augmented by one or more factors of zebrafish DNA repair systems.
KeywordReca Protein Gene Targeting Site-specific Mutagenesis Zebrafish
DepartmentUniv Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA; Univ Minnesota, Arnold & Mabel Beckman Ctr Transposon Res, St Paul, MN 55108 USA; Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China
Subject AreaBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
Indexed BySCI
WOS IDWOS:000182588700011
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Cited Times:19[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Corresponding AuthorYang, Y, Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA
Recommended Citation
GB/T 7714
Zongbin Cui,Ying Yang,Christopher D. Kaufman,et al. RecA-mediated, targeted mutagenesis in zebrafish[J]. MARINE BIOTECHNOLOGY,2003,5(2):174-184.
APA Zongbin Cui,Ying Yang,Christopher D. Kaufman,Dritan Agalliu,Perry B. Hackett,&Yang, Y, Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA.(2003).RecA-mediated, targeted mutagenesis in zebrafish.MARINE BIOTECHNOLOGY,5(2),174-184.
MLA Zongbin Cui,et al."RecA-mediated, targeted mutagenesis in zebrafish".MARINE BIOTECHNOLOGY 5.2(2003):174-184.
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