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. 2006 Jun 14;50(1-3):93–108. doi: 10.1007/s10616-006-6550-0

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

André Oumard 1, Junhua Qiao 1, Thomas Jostock 2, Jiandong Li 2, Juergen Bode 1,✉
PMCID: PMC3476001  PMID: 19003073

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.

Key words: Cassette exchange, Flexing, Floxing, Flrting, Froxing, RMCE, Site-specific recombinase, Tissue engineering

Glossary

FRT

Flp-recombinase target site

HR

homologous recombination

RMCE

recombinase-mediated cassette exchange

RT

recombinase target

SR

site-specific recombination

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