A Colony-Forming Assay for Determining the Establishment Efficiency of S/MAR-Containing Nonviral Episomal Expression Vectors
- 1Centre for Biomedical Education and Research, Institute of Cell Biology, University of Witten/Herdecke, 58453 Witten, Germany
- 2Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, 85764 Oberschleißheim, Germany
- 3Helios Medical Centre Wuppertal, Paediatrics Centre, 42117 Wuppertal, Germany
- 4Institute for Virology and Microbiology, ZBAF, University of Witten/Herdecke, 58453 Witten, Germany
Abstract
As with all eukaryotic replicons, the stable establishment of S/MAR (scaffold/matrix attached region) vectors is a stochastic event that depends on poorly understood epigenetic factors such as chromatin structure and nuclear localization. Establishment efficiency describes the percentage of cells in which a particular S/MAR vector is stably retained as an episome after an initial selection period. Expected establishment efficiency for S/MAR vectors is 1–5%. This article describes a colony-forming assay that may be used either to determine establishment efficiency or to generate single cell clones.
Footnotes
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↵5 Correspondence: claudia.hagedorn{at}uni-wh.de
- © 2012 Cold Spring Harbor Laboratory Press
