Restriction enzyme mediated integration

Restriction enzyme mediated integration is a technique for integrating DNA into the genome sites that have been generated by the same restriction enzyme used for the DNA linearisation. The plasmid integration occurs at the corresponding sites in the genome, often by regenerating the recognition sites by same the restriction enzyme used for plasmid linearisation.

Mechanism

The specific restriction enzyme cleaves the genomic DNA at random points, and generates recognition sites. The DNA fragment to be inserted is linearised using the said same restriction enzyme and the mix injected into the cell followed by a successful insertion of a DNA fragment.

Applications

The REMI method has been used to generate genetically modified organisms including the Saccharomyces cerevisiae, Dictyostelium discoideum and Xenopus laevis.

Genome engineering

REMI has been used for large-scale transgenesis in X. laevis embryos in order to study various signaling pathways including the FGF and the Thyroid system.