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
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]
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]
^Kessler, C.; Manta, V. (1990-08-16). "Specificity of restriction endonucleases and DNA modification methyltransferases a review (Edition 3)". Gene. 92 (1–2): 1–248. doi:10.1016/0378-1119(90)90486-b. ISSN0378-1119. PMID2172084.
^ abKroll, K. L.; Amaya, E. (1996-10-01). "Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation". Development. 122 (10): 3173–3183. doi:10.1242/dev.122.10.3173. ISSN0950-1991. PMID8898230. S2CID20349983.
