Site-specific and directional gene replacement mediated by Cre recombinase.
A novel method for the site-specific introduction of genes into eukaryotic cells using the prokaryotic Cre-LoxP recombination system is presented. Cre recombinase catalyzes recombination between two LoxP sites or between two mutant LoxP 511 sites. However, recombination is not catalyzed between a LoxP and a LoxP 511 site. We now demonstrate that it is possible to catalyze accurate exchange between two DNA segments each flanked by a LoxP and a LoxP 511 site. In the example presented, expression of the Cre recombinase resulted in the replacement of a murine IgA constant region gene with a LoxP site at the 5' end and a LoxP 511 site at the 3' end by a human IgA constant region gene flanked by the same wild type and mutant LoxP sites. This method provides a novel approach for the site-specific substitution of specific genes.
Trinh
K R
KR
Department of Microbiology and Molecular Genetics, University of California, Los Angeles, CA 90095-1747, USA.
Morrison
S L
SL
D000818:Animals; D017101:Bacteriophage P1 / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis / Q000235:genetics; D019426:Integrases / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014764:Viral Proteins
D007070:Immunoglobulin A; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases