Unidirectional Cre-mediated genetic inversion in mice using the mutant loxP pair lox66/lox71.
The Cre/loxP recombination system is a commonly used tool to alter the mouse genome in a conditional manner by deletion or inversion of loxP-flanked DNA segments. While Cre-mediated deletion is essentially unidirectional, inversion is reversible and therefore does not allow the stable alteration of gene function in cells that constitutively express Cre. Site-directed mutagenesis yielded a pair of asymmetric loxP sites (lox66 and lox71) that display a favorable forward reaction equilibrium. Here, we demonstrate that lox66/lox71 mediates efficient and predominantly unidirectional inversion of a switch substrate targeted to the mouse genome in combination with either inducible or cell type-specific cre-transgenes in vivo.
Oberdoerffer
Philipp
P
The CBR Institute for Biomedical Research, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
Otipoby
Kevin L
KL
Maruyama
Mitsuo
M
Rajewsky
Klaus
K
D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D015139:Blotting, Southern; D002460:Cell Line; D004247:DNA / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D015321:Gene Rearrangement / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D011995:Recombination, Genetic; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D007143:Immunoglobulin Heavy Chains; D014764:Viral Proteins; D004247:DNA; C045073:Cre recombinase; D019426:Integrases