title: A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy.
abstract: 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.
authors.affiliation: Chromos Molecular Systems, Inc., 8081 Lougheed Highway, Burnaby, BC, Canada V5A 1W9.
authors.forename: Michael
authors.initials: M
authors.lastname: Lindenbaum
authors.forename: Ed
authors.initials: E
authors.lastname: Perkins
authors.forename: Erika
authors.initials: E
authors.lastname: Csonka
authors.forename: Elena
authors.initials: E
authors.lastname: Fleming
authors.forename: Lisa
authors.initials: L
authors.lastname: Garcia
authors.forename: Amy
authors.initials: A
authors.lastname: Greene
authors.forename: Lindsay
authors.initials: L
authors.lastname: Gung
authors.forename: Gyula
authors.initials: G
authors.lastname: Hadlaczky
authors.forename: Edmond
authors.initials: E
authors.lastname: Lee
authors.forename: Josephine
authors.initials: J
authors.lastname: Leung
authors.forename: Neil
authors.initials: N
authors.lastname: MacDonald
authors.forename: Alexisann
authors.initials: A
authors.lastname: Maxwell
authors.forename: Kathleen
authors.initials: K
authors.lastname: Mills
authors.forename: Diane
authors.initials: D
authors.lastname: Monteith
authors.forename: Carl F
authors.initials: CF
authors.lastname: Perez
authors.forename: Joan
authors.initials: J
authors.lastname: Shellard
authors.forename: Sandy
authors.initials: S
authors.lastname: Stewart
authors.forename: Tom
authors.initials: T
authors.lastname: Stodola
authors.forename: Dana
authors.initials: D
authors.lastname: Vandenborre
authors.forename: Sandy
authors.initials: S
authors.lastname: Vanderbyl
authors.forename: Harry C
authors.initials: HC
authors.lastname: Ledebur
mesh_terms: 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
chemical_list: D011994:Recombinant Proteins; D004921:Erythropoietin; D019426:Integrases