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

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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.

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Abbreviations

FRT:

Flp-recombinase target site

HR:

homologous recombination

RMCE:

recombinase-mediated cassette exchange

RT:

recombinase target

SR:

site-specific recombination

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Authors and Affiliations

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

    André Oumard, Junhua Qiao & Juergen Bode

  2. Technical University of Braunschweig, Institute of Biochemistry and Biotechnology, Spielmannstr. 7, D-38106, Braunschweig, Germany

    Thomas Jostock & Jiandong Li

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  1. André Oumard
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  2. Junhua Qiao
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  3. Thomas Jostock
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  4. Jiandong Li
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  5. Juergen Bode
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Correspondence to Juergen Bode.

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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 50, 93–108 (2006). https://doi.org/10.1007/s10616-006-6550-0

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