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Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.
Qiao, Junhua Oumard, André Wegloehner, Wolfgang Bode, Juergen
Qiao, Junhua
Oumard, André
Wegloehner, Wolfgang
Bode, Juergen
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Issue Date
2009-07-24
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Abstract
Site-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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Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties. 2009, 390 (4):579-94 J. Mol. Biol.
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en
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1089-8638