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dc.contributor.authorQiao, Junhua
dc.contributor.authorOumard, André
dc.contributor.authorWegloehner, Wolfgang
dc.contributor.authorBode, Juergen
dc.date.accessioned2009-08-07T09:05:59Z
dc.date.available2009-08-07T09:05:59Z
dc.date.issued2009-07-24
dc.identifier.citationNovel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties. 2009, 390 (4):579-94 J. Mol. Biol.en
dc.identifier.issn1089-8638
dc.identifier.pmid19447116
dc.identifier.doi10.1016/j.jmb.2009.05.012
dc.identifier.urihttp://hdl.handle.net/10033/76653
dc.description.abstractSite-specific recombinases have revolutionized the systematic generation of transgenic cell lines and embryonic stem cells/animals and will ultimately also reveal their potential in the genetic modification of induced pluripotent stem cells. Introduced in 1994, our Flp recombinase-mediated cassette exchange strategy permits the exchange of a target cassette for a cassette with the gene of interest, introduced as a part of an exchange vector. The process is "clean" in the sense that it does not co-introduce prokaryotic vector parts; neither does it leave behind a selection marker. Stringent selection principles provide master cell lines permitting subsequent recombinase-mediated cassette exchange cycles in the absence of a drug selection and with a considerable efficiency (approximately 10%). Exemplified by Chinese hamster ovary cells, the strategy proves to be successful even for cell lines with an unstable genotype.
dc.language.isoenen
dc.titleNovel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.en
dc.typeArticleen
dc.contributor.departmentDepartment of Molecular Biotechnology/Epigenetic Regulation, Helmholtz Centre for Infection Research, Braunschweig, Germany.en
dc.identifier.journalJournal of molecular biologyen
refterms.dateFOA2018-06-13T00:22:50Z
html.description.abstractSite-specific recombinases have revolutionized the systematic generation of transgenic cell lines and embryonic stem cells/animals and will ultimately also reveal their potential in the genetic modification of induced pluripotent stem cells. Introduced in 1994, our Flp recombinase-mediated cassette exchange strategy permits the exchange of a target cassette for a cassette with the gene of interest, introduced as a part of an exchange vector. The process is "clean" in the sense that it does not co-introduce prokaryotic vector parts; neither does it leave behind a selection marker. Stringent selection principles provide master cell lines permitting subsequent recombinase-mediated cassette exchange cycles in the absence of a drug selection and with a considerable efficiency (approximately 10%). Exemplified by Chinese hamster ovary cells, the strategy proves to be successful even for cell lines with an unstable genotype.


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