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Oumard A, Qiao J, Jostock T, et al. Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology. Cytotechnology. 2006;50(1):93-108doi: 10.1007/s10616-006-6550-0.
Oumard, A., Qiao, J., Jostock, T., Li, J., & Bode, J. (2006). Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology. Cytotechnology, 50(1), 93-108. https://doi.org/10.1007/s10616-006-6550-0
Oumard, André, et al. "Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology." Cytotechnology vol. 50,1 (2006): 93-108. doi: https://doi.org/10.1007/s10616-006-6550-0
Oumard A, Qiao J, Jostock T, Li J, Bode J. Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology. Cytotechnology. 2006 Mar;50(1):93-108. doi: 10.1007/s10616-006-6550-0. Epub 2006 Jun 14. PMID: 19003073; PMCID: PMC3476001.
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Cytotechnology. 2006 Mar;50(1):93-108. doi: 10.1007/s10616-006-6550-0. Epub 2006 Jun 14.

Recommended Method for Chromosome Exploitation: RMCE-based Cassette-exchange Systems in Animal Cell Biotechnology.

Cytotechnology

André Oumard, Junhua Qiao, Thomas Jostock, Jiandong Li, Juergen Bode

Affiliations

  1. German Research Center for Biotechnology (GBF), RDIF/Epigenetic Regulation, Mascheroder Weg 1, D-38124, Braunschweig, Germany.

PMID: 19003073 PMCID: PMC3476001 DOI: 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 PhiC31, 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 PhiC31. 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.

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