FLP-mediated site-specific recombination in microinjected murine zygotes.
The FLP recombinase of yeast catalyses site-specific recombination between repeated FLP recombinase target (FRT) elements in yeast and in heterologous systems (Escherichia coli, Drosophila, mosquito and cultured mammalian cells). In this report, it is shown that transient FLP recombinase expression can recombine and activate an extrachromosomal silent reporter gene following coinjection into fertilized one-cell mouse eggs. Furthermore, it is demonstrated that introduction of a FLP-recombinase expression vector into transgenic one-cell fertilized mouse eggs induces a recombination event at a chromosomal FRT target locus. The resulting event occurred at the one-cell stage and deleted a chromosomal tandem array of a FRT containing lacZ expression cassette down to one or two copies. These results demonstrate that the FLP recombinase can be utilized to manipulate the genome of transgenic animals and suggest that FLP recombinase-mediated plasmid-to-chromosome targeting is feasible in microinjected eggs.
Ludwig
D L
DL
Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati Medical Center, OH 45267-0524, USA.
Stringer
J R
JR
Wight
D C
DC
Doetschman
H C
HC
Duffy
J J
JJ
D000818:Animals; D002875:Chromosomes; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008845:Microinjections; D011247:Pregnancy; D011995:Recombination, Genetic; D015053:Zygote
D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase