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Mol. Cell. Biol., Apr 1995, 1968-1973, Vol 15, No. 4
Copyright © 1995, American Society for Microbiology

Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae

A Choulika, A Perrin, B Dujon and JF Nicolas
Unite de Biologie moleculaire du Developpement, Institut Pasteur, Universite Pierre et Marie Curie, Paris, France.

The mitochondrial intron-encoded endonuclease I-SceI of Saccharomyces cerevisiae has an 18-bp recognition sequence and, therefore, has a very low probability of cutting DNA, even within large genomes. We demonstrate that double-strand breaks can be initiated by the I-SceI endonuclease at a predetermined location in the mouse genome and that the breaks can be repaired with a donor molecule homologous regions flanking the breaks. This induced homologous recombination is approximately 2 orders of magnitude more frequent than spontaneous homologous recombination and at least 10 times more frequent than random integration near an active promoter. As a consequence of induced homologous recombination, a heterologous novel sequence can be inserted at the site of the break. This recombination can occur at a variety of chromosomal targets in differentiated and multipotential cells. These results demonstrate homologous recombination involving chromosomal DNA by the double-strand break repair mechanism in mammals and show the usefulness of very rare cutter endonucleases, such as I-SceI, for designing genome rearrangements.


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