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A genetic screen identifies novel non-compatible loxP sites

2002; Oxford University Press; Volume: 30; Issue: 14 Linguagem: Inglês

10.1093/nar/gkf421

1362-4962

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

Advanced biosensing and bioanalysis techniques

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.