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Nucleic Acids Research, 2002, Vol. 30, No. 14 3067-3077
© 2002 Oxford University Press

A genetic screen identifies novel non-compatible loxP sites

Stephen J. Langer, A. Paiman Ghafoori, Marshall Byrd and Leslie Leinwand*

Department of Molecular, Cellular and Developmental Biology, CB347, University of Colorado, Boulder, CO 80309-0347, USA

*To whom correspondence should be addressed. Tel: +1 303 492 7606; Fax: +1 303 492 8907; Email: leinwand{at}stripe.colorado.edu
Present addresses:
A. Paiman Ghafoori, Harvard Medical School, Boston, MA 02115, USA
Marshall Byrd, Department of Molecular Virology and Microbiology, Baylor College, Houston, TX 77030, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

The ability of the Cre/lox system to make precise genomic modifications is a tremendous accomplishment. However, recombination between cis-linked heterospecific lox sites limits the use of Cre- mediated exchange of DNA to systems where genetic selection can be applied. To circumvent this problem we carried out a genetic screen designed to identify novel mutant spacer-containing lox sites displaying enhanced incompatibility with the canonical loxP site. One of the mutant sites recovered appears to be completely stable in HEK293 cells constitutively expressing Cre recombinase and supports recombinase-mediated cassette exchange (RMCE) in bacteria and mammalian cell culture. By preventing undesirable recombination, these novel lox sites could improve the efficiency of in vivo gene transfer.


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