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AssigneeE. I. Du Pont de Nemours and Company
InventorsSaverio Carl Falco, Zhongsen Li
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A targeting method is described that allows precise cassette replacement at a previously characterized genetic locus. A target DNA construct containing a pair of incompatible FRT sites flanking a target cassette was introduced into soybean by regular biolistic transformation. Transgenic events containing a single complete copy of the target site were then selected and retransformed with a donor DNA construct containing the identical pair of incompatible FRT sites flanking a donor cassette. Precise DNA cassette exchange happened between the target cassette and the donor cassette via recombinase mediated cassette exchange (RMCE) so that the donor cassette was introduced at the exact genomic site previously occupied by the target cassette. Through repeated RMCE using additional incompatible FRT sites, multiple groups of transgenes can be stacked at the same genomic locus.
A soybean cell, plant or seed having stably incorporated in its genome a transfer cassette genetically linked to a chromosomal region comprising SEQ ID NO:80 or SEQ ID NO:84, wherein the transfer cassette comprises at least two non-identical recombination sites, wherein the transfer cassette further comprises a polynucleotide encoding a selectable marker protein-coding sequence bounded by a first recombination site and a second non-identical recombination site.
A method for stacking of multiple expression cassettes of interest into a specific chromosomal site in a soybean genome, said method comprising:
Dependencies are inferred from the claim text.