A colony-forming assay for determining the establishment efficiency of S/MAR-containing nonviral episomal expression vectors.
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.
Centre for Biomedical Education and Research, Institute of Cell Biology, University of Witten/Herdecke, 58453 Witten, Germany.
Claudia
C
Hagedorn
Armin
A
Baiker
Jan
J
Postberg
Anja
A
Ehrhardt
Hans J
HJ
Lipps
D000818:Animals; D016466:CHO Cells; D003114:Colony-Forming Units Assay / Q000379:methods*; D006224:Cricetinae; D004261:DNA Replication; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D042822:Genomic Instability; D010957:Plasmids; D014170:Transformation, Genetic