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Nucleic Acids Research 2004 32(21):e172; doi:10.1093/nar/gnh169
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Published online 7 December 2004

Nucleic Acids Research, Vol. 32 No. 21 © Oxford University Press 2004; all rights reserved

A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy

Michael Lindenbaum, Ed Perkins, Erika Csonka1, Elena Fleming, Lisa Garcia, Amy Greene, Lindsay Gung, Gyula Hadlaczky1, Edmond Lee, Josephine Leung, Neil MacDonald, Alexisann Maxwell, Kathleen Mills, Diane Monteith, Carl F. Perez*, Joan Shellard, Sandy Stewart, Tom Stodola, Dana Vandenborre, Sandy Vanderbyl and Harry C. Ledebur, Jr

Chromos Molecular Systems, Inc., 8081 Lougheed Highway, Burnaby, BC, Canada V5A 1W9 and 1 Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Temesvári krt. 62, PO Box 521, H-6701 Szeged, Hungary

* To whom correspondence should be addressed. Tel: +1 604 415 7100; Fax: +1 604 415 7151; Email: cperez{at}chromos.com
Present addresses: Michael Lindenbaum, Agrisoma Biosciences, Inc., 110 Gymnasium Place, Saskatoon, Saskatchewan, Canada S7N 0W9
Ed Perkins and Amy Greene, Department of Biochemistry and Molecular Biology, University of Minnesota School of Medicine, Duluth Campus, 1035 University Drive, Duluth, MN 55812-3031, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Received August 21, 2004; Revised and Accepted November 15, 2004

Mammalian artificial chromosomes (MACs) provide a means to introduce large payloads of genetic information into the cell in an autonomously replicating, non-integrating format. Unique among MACs, the mammalian satellite DNA-based Artificial Chromosome Expression (ACE) can be reproducibly generated de novo in cell lines of different species and readily purified from the host cells' chromosomes. Purified mammalian ACEs can then be re-introduced into a variety of recipient cell lines where they have been stably maintained for extended periods in the absence of selective pressure. In order to extend the utility of ACEs, we have established the ACE System, a versatile and flexible platform for the reliable engineering of ACEs. The ACE System includes a Platform ACE, containing >50 recombination acceptor sites, that can carry single or multiple copies of genes of interest using specially designed targeting vectors (ATV) and a site-specific integrase (ACE Integrase). Using this approach, specific loading of one or two gene targets has been achieved in LMTK– and CHO cells. The use of the ACE System for biological engineering of eukaryotic cells, including mammalian cells, with applications in biopharmaceutical production, transgenesis and gene-based cell therapy is discussed.


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