Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology
- 1 March 2006
- journal article
- research article
- Published by Springer Science and Business Media LLC in Cytotechnology
- Vol. 50 (1-3), 93-108
- https://doi.org/10.1007/s10616-006-6550-0
Abstract
The availability of site-specific recombinases has revolutionized the rational construction of cell lines with predictable properties. Early efforts were directed to providing pre-characterized genomic loci with a single recombinase target site that served as an address for the integration of vectors carrying a compatible tag. Efficient procedures of this type had to await recombinases like ΦC31, which recombine attP and attB target sites in a one-way reaction — at least in the cellular environment of the higher eukaryotic cell. Still these procedures lead to the co-introduction of prokaryotic vector sequences that are known to cause epigenetic silencing. This review illuminates the actual status of the more advanced recombinase-mediated cassette exchange (RMCE) techniques that have been developed for the major members of site-specific recombinases (SR), Flp, Cre and ΦC31. In RMCE the genomic address consists of a set of heterospecific recombinase target (RT-) sites permitting the exchange of the intervening sequence for the gene of interest (GOI), as part of a similar cassette. This process locks the GOI in place and it is ‘clean’ in the sense that it does not co-introduce prokaryotic vector parts nor does it leave behind a selection marker.Keywords
This publication has 48 references indexed in Scilit:
- A New Generation of Retroviral Producer Cells: Predictable and Stable Virus Production by Flp-Mediated Site-Specific Integration of Retroviral VectorsMolecular Therapy, 2006
- Correlations between Scaffold/Matrix Attachment Region (S/MAR) Binding Activity and DNA Duplex Destabilization EnergyJournal of Molecular Biology, 2006
- Mutant Lrp1 Knock-In Mice Generated by Recombinase-Mediated Cassette Exchange Reveal Differential Importance of the NPXY Motifs in the Intracellular Domain of LRP1 for Normal Fetal DevelopmentMolecular and Cellular Biology, 2006
- Increased Maintenance and Persistence of Transgenes by Excision of Expression Cassettes from Plasmid SequencesIn VivoHuman Gene Therapy, 2005
- Performance of Genomic Bordering Elements at Predefined Genomic LociMolecular and Cellular Biology, 2005
- The dynamic response of upstream DNA to transcription-generated torsional stressNature Structural & Molecular Biology, 2004
- Stable and strictly controlled expression of LTR-flanked autoregulated expression cassettes upon adenoviral transferBiochemical and Biophysical Research Communications, 2004
- Talking about a Revolution: The Impact of Site-Specific Recombinases on Genetic Analyses in MiceDevelopmental Cell, 2004
- Elk-1 knock-out mice engineered by Flp recombinase-mediated cassette exchangegenesis, 2004
- Use of Mutated FLP Recognition Target (FRT) Sites for the Exchange of Expression Cassettes at Defined Chromosomal LociBiochemistry, 1994