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Nucleic Acids Research, 2003, Vol. 31, No. 21 e131
© 2003 Oxford University Press

Regulation of Cre recombinase by ligand-induced complementation of inactive fragments

Nicolas Jullien, François Sampieri1, Alain Enjalbert and Jean-Paul Herman*

ICNE-UMR 6544 and 1 UMR 6560 CNRS-Université de la Méditerranée, IFR Jean-Roche, Faculté de Médecine Nord, 13916 Marseille Cedex 20, France

*To whom correspondence should be addressed. Tel: +33 491 69 87 15; Fax: +33 491 69 89 20; Email: herman.jp{at}jean-roche.univ-mrs.fr

Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. To overcome this, we have developed DiCre, a regulatable fragment complementation system for Cre. The enzyme was split into two moieties that were fused to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12–rapamycin-associated protein), respectively. These can be efficiently heterodimerized by rapamycin. Several variants, based on splitting Cre at different sites and using different linker peptides, were tested in an indicator cell line. The fusion proteins, taken separately, had no recombinase activity. Stable transformants, co-expressing complementing fragments based on splitting Cre between Asn59 and Asn60, displayed low background activity affecting 0.05–0.4% of the cells. Rapamycin induced a rapid recombination, reaching 100% by 48–72 h, with an EC50 of 0.02 nM. Thus, ligand-induced dimerization can efficiently regulate Cre, and should be useful to achieve a tight temporal control of its activity, such as in the case of the creation of conditional knock-out animals.


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