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Nucleic Acids Research, 2002, Vol. 30, No. 19 e103
© 2002 Oxford University Press

Site-directed integration of the cre gene mediated by Cre recombinase using a combination of mutant lox sites

Kimi Araki*, Masatake Araki1 and Ken-ichi Yamamura

Department of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University, Kuhonnji 4-24-1, Kumamoto 862-0976, Japan and 1 Gene Technology Center, Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811, Japan

*To whom correspondence should be addressed. Tel: +81 96 373 6598; Fax: +81 96 373 6599; Email: arakimi{at}gpo.kumamoto-u.ac.jp

The Cre-lox system is an important tool for genetic manipulation. To promote integrative reactions, two strategies using mutant lox sites have been developed. One is the left element/right element (LE/RE)-mutant strategy and the other is the cassette exchange strategy using heterospecific lox sites such as lox511 or lox2272. We compared the recombination efficiencies using these mutant lox sites in embryonic stem (ES) cells, and found that the combination of the LE/RE mutant and lox2272 showed high recombination efficiency and stability of the recombined structure. Taking advantage of this stability, we successfully integrated the cre gene into the mutant lox sites by Cre-mediated recombination. Germ line chimeric mice were produced from the cre-integrated ES cell clones, and Cre-expressing mouse lines were established. The inserted cre gene was stably maintained through the generations. This cre knock-in system using the LE/RE-lox2272 combination should be useful for the production of various cre mice for gene targeting or gene trapping.


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