Patent ID: 8148607
Filing Date: 2012-04-03
Classification: C12N

Abstract:
1. A method for exchanging a target DNA sequence in the genome of a plant for a DNA sequence of interest comprising: a. inducing a first double stranded DNA break at a preselected site in the genome of a cell of a plant, said preselected site being located within said target DNA sequence or in the vicinity of said target DNA sequence; b. introducing a repair DNA molecule into said plant cell, said DNA molecule comprising c. selecting a population of plant cells comprising said selectable or screenable marker; d. selecting a plant cell wherein said selectable or screenable marker has been introduced by homologous recombination through said flanking DNA regions and regenerating a plant from said plant cell; e. crossing said regenerated plant or a progeny plant thereof comprising said selectable marker gene with a plant comprising a DSBI enzyme encoding chimeric gene, said chimeric gene comprising the following operably linked DNA segments: f. selecting a progeny plant (F1-plant) comprising said selectable or screenable marker gene and said DSBI enzyme encoding chimeric gene; g. crossing said progeny plant with another plant whereby said progeny plant is used as pollen donor; h. selecting a population of progeny plants (F2-population) which comprises said DSBI enzyme encoding chimeric gene; and i. selecting a progeny plant wherein said selectable or screenable marker gene is deleted by homologous recombination between said one of the flanking DNA regions and said another copy of at least part of said one of the flanking DNA regions immediately adjacent to the target DNA sequence, located in direct repeat.