Nucleic Acids Research, Vol 24, Issue 19 3707-3713, Copyright © 1996 by Oxford University Press
G Camenisch, M Gruber, G Donoho, P Van Sloun, RH Wenger and M Gassmann
We describe the ability of novel episomally maintained vectors to
efficiently promote gene expression in embryonic stem (ES) cells as well as
in established mouse cell lines. Extrachromosomal maintenance of our
vectors is based on the presence of polyoma virus DNA sequences, including
the origin of replication harboring a mutant enhancer (PyF101), and a
modified version of the polyoma early region (LT20) encoding the large T
antigen only. Reporter gene expression from such extrachromosomally
replicating vectors was approximately 10-fold higher than expression from
replication-incompetent control plasmids. After transfection of different
ES cell lines, the polyoma virus-derived plasmid variant pMGD20neo (7.2 kb)
was maintained episomally in 16% of the G418-resistant clones. No
chromosomal integration of pMGD20neo vector DNA was detected in ES cells
that contained episomal vector DNA even after long term passage. The
vector's replication ability was not altered after insertion of up to 10 kb
hprt gene fragments. Besides undifferentiated ES cells, the polyoma-based
vectors were also maintained extrachromosomally in differentiating ES cells
and embryoid bodies as well as in established mouse cell lines.
ARTICLES
A polyoma-based episomal vector efficiently expresses exogenous genes in mouse embryonic stem cells
Institute of Physiology, University of Zurich, Switzerland.
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