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

Détails

ID Serval
serval:BIB_46D974BC2B64
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.
Périodique
Journal of Molecular Biology
Auteur(s)
Qiao J., Oumard A., Wegloehner W., Bode J.
ISSN
1089-8638[electronic], 0022-2836[linking]
Statut éditorial
Publié
Date de publication
2009
Peer-reviewed
Oui
Volume
390
Numéro
4
Pages
579-594
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
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.
Mots-clé
Animals, CHO Cells, Clone Cells, Cloning, Molecular, Cricetinae, Cricetulus, DNA Nucleotidyltransferases/genetics, Flow Cytometry, Gene Targeting, Green Fluorescent Proteins/genetics, Recombination, Genetic, Transgenes
Pubmed
Création de la notice
16/06/2010 12:11
Dernière modification de la notice
03/03/2018 16:48
Données d'usage