Restriction enzyme mediated integration
Restriction enzyme mediated integration (abbreviated as REMI) is a technique for integrating DNA (linearised plasmid) into the genome sites that have been generated by the same restriction enzyme used for the DNA linearisation.[1] 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
[edit | edit source]The specific restriction enzyme cleaves the genomic DNA at random points, and generates recognition sites.[2] 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.[3]
Applications
[edit | edit source]The REMI method has been used to generate genetically modified organisms including the Saccharomyces cerevisiae, Dictyostelium discoideum and Xenopus laevis.[1][4][5]
Genome engineering
[edit | edit source]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.[3][6]
References
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