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Publication : Cre reconstitution allows for DNA recombination selectively in dual-marker-expressing cells in transgenic mice.

First Author  Xu Y Year  2007
Journal  Nucleic Acids Res Volume  35
Issue  19 Pages  e126
PubMed ID  17893102 Mgi Jnum  J:172178
Mgi Id  MGI:5004791 Doi  10.1093/nar/gkm559
Citation  Xu Y, et al. (2007) Cre reconstitution allows for DNA recombination selectively in dual-marker-expressing cells in transgenic mice. Nucleic Acids Res 35(19):e126
abstractText  Cre/LoxP-based DNA recombination has been used to introduce desired DNA rearrangements in various organisms, having for example, greatly assisted genetic analyses in mice. For most applications, single gene promoters are used to drive Cre production for conditional gene activation/inactivation or lineage-tracing experiments. Such a manipulation introduces Cre in all cells in which the utilized promoter is active. To overcome the limited selectivity of single promoters for cell-type-specific recombination, we have explored the 'dual promoter combinatorial control' of Cre activity, so that Cre activity could be restricted to cells that express dual protein markers. We efficiently reconstituted Cre activity from two modified, inactive Cre fragments. Cre re-association was greatly enhanced by fusing the Cre fragments separately to peptides that can form a tight antiparallel leucine zipper. The co-expressed Cre fusion fragments showed substantial activity in cultured cells. As proof of principle of the utility of this technique in vivo for manipulating genes specifically in dual-marker-positive cells, we expressed each inactive Cre fragments in transgenic mice via individual promoters. Result showed the effective reconstitution of Cre activates LoxP recombination in the co-expressing cells.
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