A genetic screen identifies novel non-compatible loxP sites | Zendy

Stephen J. Langer | Zendy; A. Paiman Ghafoori | Zendy; Marshall Byrd | Zendy; Leslie A. Leinwand | Zendy

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A genetic screen identifies novel non-compatible loxP sites

Author(s) -

Stephen J. Langer,

A. Paiman Ghafoori,

Marshall Byrd,

Leslie A. Leinwand

Publication year - 2002

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkf421

Subject(s) - biology , cre recombinase , recombinase , cre lox recombination , mutant , site specific recombination , genetics , flp frt recombination , dna , genetic screen , hek 293 cells , mutation , gene , gene targeting , computational biology , recombination , genetic recombination , transgene , genetically modified mouse

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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