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

2002; Oxford University Press; Volume: 30; Issue: 19 Linguagem: Inglês

10.1093/nar/gnf102

1362-4962

Koji Araki,

RNA Interference and Gene Delivery

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 lox 511 or lox 2272. 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 lox 2272 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‐ lox 2272 combination should be useful for the production of various cre mice for gene targeting or gene trapping.