A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy.
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.
Lindenbaum
Michael
M
Chromos Molecular Systems, Inc., 8081 Lougheed Highway, Burnaby, BC, Canada V5A 1W9.
Perkins
Ed
E
Csonka
Erika
E
Fleming
Elena
E
Garcia
Lisa
L
Greene
Amy
A
Gung
Lindsay
L
Hadlaczky
Gyula
G
Lee
Edmond
E
Leung
Josephine
J
MacDonald
Neil
N
Maxwell
Alexisann
A
Mills
Kathleen
K
Monteith
Diane
D
Perez
Carl F
CF
Shellard
Joan
J
Stewart
Sandy
S
Stodola
Tom
T
Vandenborre
Dana
D
Vanderbyl
Sandy
S
Ledebur
Harry C
HC
D000818:Animals; D030801:Animals, Genetically Modified; D016466:CHO Cells; D002460:Cell Line; D022221:Chromosomes, Artificial, Mammalian; D006224:Cricetinae; D003412:Cricetulus; D004345:Drug Industry; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy; D006801:Humans; D019426:Integrases / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins; D004921:Erythropoietin; D019426:Integrases