Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.

Details

Serval ID
serval:BIB_46D974BC2B64
Type
Article: article from journal or magazin.
Collection
Publications
Title
Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.
Journal
Journal of Molecular Biology
Author(s)
Qiao J., Oumard A., Wegloehner W., Bode J.
ISSN
1089-8638[electronic], 0022-2836[linking]
Publication state
Published
Issued date
2009
Peer-reviewed
Oui
Volume
390
Number
4
Pages
579-594
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
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.
Keywords
Animals, CHO Cells, Clone Cells, Cloning, Molecular, Cricetinae, Cricetulus, DNA Nucleotidyltransferases/genetics, Flow Cytometry, Gene Targeting, Green Fluorescent Proteins/genetics, Recombination, Genetic, Transgenes
Pubmed
Create date
16/06/2010 11:11
Last modification date
20/08/2019 13:52
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