Homologous recombination: a basis for targeted genome optimization in crop species such as maize

被引:62
|
作者
D'Halluin, Kathleen [1 ]
Vanderstraeten, Chantal [1 ]
Stals, Ellen [1 ]
Cornelissen, Marc [1 ]
Ruiter, Rene [1 ]
机构
[1] Bayer BioSci NV, B-9052 Ghent, Belgium
关键词
double-strand break repair; gene targeting; homologous recombination; maize; transformation;
D O I
10.1111/j.1467-7652.2007.00305.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In an attempt to understand the feasibility of future targeted genome optimization in agronomic crops, we tested the efficiency of homologous recombination-mediated sequence insertion upon induction of a targeted DNA double-strand break at the desired integration site in maize. By the development of an efficient tissue culture protocol, and with the use of an I-SceI gene optimized for expression in maize, large numbers of precisely engineered maize events were produced in which DNA integration occurred very accurately. In a subset of events examined in detail, no additional deletions and/or insertions of short filler DNA at the integration site were observed. In 30%-40% of the recovered events, no traces of random insertions were observed. This was true for DNA delivery by both Agrobacterium and particle bombardment. These data suggest that targeted double-strand break-induced homologous recombination is a superior method to generate specific desired changes in the maize genome, and suggest targeted genome optimization of agronomic crops to be feasible.
引用
收藏
页码:93 / 102
页数:10
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