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.