|Srivastava, Vibha - USDA/UCB PGEC|
Submitted to: Patent Application
Publication Type: Other
Publication Acceptance Date: September 5, 2000
Publication Date: September 5, 2000
Interpretive Summary: This patent describes using a set of inverted recombination sites to flank the introduced DNA, such that excess copies of the introduced DNA can be reduced to a single copy state by stably or transiently produced recombinase proteins.
Technical Abstract: The present invention provides methods for producing a transgenic cell having a stably integrated, single copy of an exogenous polynucleotide sequence. The method, which resolves repeated insertions of the introduced polynucleotide sequence into a single copy, involves introducing into a genetic locus of a cell a polynucleotide sequence flanked on each end by a recombination site. The recombination sites are oriented such that contact with a recombinase would not result in deletion of the polynucleotide sequence from the construct. The multiple, tandem copies of the introduced polynucleotide locus are then contacted with a recombinase that catalyzes recombination among the recombination sites. As a result of this method, the multiple, tandem copies are resolved to a single copy.