Use of mutated FLP recognition target (FRT) sites for the exchange of expression cassettes at defined chromosomal loci.
Using the FLP/FRT system for site-specific recombination and the wild-type recognition site (FRT) in conjunction with certain mutant FRT sites, it becomes possible to provoke, with high yield, a double-reciprocal crossover event in cultured mammalian cells. It is demonstrated that this technology enables a targeting of expression cassettes to appropriate chromosomal reference sites in the recipient cell to improve the concepts of reverse genetics. The design of mutant FRT sites promoting such a process will be delineated. Our results show that the five spacer mutations tested are functional as the wild type but differ in the extent of their cross-recombination, which has to be minimized for their simultaneous usage.
Schlake
T
T
GBF, Gesellschaft für Biotechnologische Forschung mbH, Braunschweig-Stöckheim, Germany.
Bode
J
J
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D007668:Kidney; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection; D014166:Transferases; D001616:beta-Galactosidase / Q000235:genetics
D004247:DNA; D014166:Transferases; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase; D001616:beta-Galactosidase