Open AccessHomologous recombination: a basis for targeted genome optimization in crop species such as maize
Author(s) -
D’Halluin Kathleen,
Vanderstraeten Chantal,
Stals Ellen,
Cornelissen Marc,
Ruiter Rene
Publication year - 2008
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/j.1467-7652.2007.00305.x
Subject(s) - biology , homologous recombination , genome , dna , recombination , genetics , gene , homologous chromosome , computational biology , dna sequencing
Summary 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‐ Sce I 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.