title: Enhanced protein production by engineered zinc finger proteins.
abstract: Increasing the yield of therapeutic proteins from mammalian production cell lines reduces costs and decreases the time to market. To this end, we engineered a zinc finger protein transcription factor (ZFP TF) that binds a DNA sequence within the promoter driving transgene expression. This ZFP TF enabled >100% increase in protein yield from CHO cells in transient, stable, and fermentor production run settings. Expression vectors engineered to carry up to 10 ZFP binding sites further enhanced ZFP-mediated increases in protein production up to approximately 500%. The multimerized ZFP binding sites function independently of the promoter, and therefore across vector platforms. CHO cell lines stably expressing ZFP TFs demonstrated growth characteristics similar to parental cell lines. ZFP TF expression and gains in protein production were stable over >30 generations in the absence of antibiotic selection. Our results demonstrate that ZFP TFs can rapidly and stably increase protein production in mammalian cells.
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Richmond, CA 94804, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Reik
authors.forename: Yuanyue
authors.initials: Y
authors.lastname: Zhou
authors.forename: Trevor N
authors.initials: TN
authors.lastname: Collingwood
authors.forename: Lyndon
authors.initials: L
authors.lastname: Warfe
authors.forename: Victor
authors.initials: V
authors.lastname: Bartsevich
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Kong
authors.forename: Karla A
authors.initials: KA
authors.lastname: Henning
authors.forename: Barrett K
authors.initials: BK
authors.lastname: Fallentine
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Zhong
authors.forename: Yann
authors.initials: Y
authors.lastname: Jouvenot
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Jamieson
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: Casey C
authors.initials: CC
authors.lastname: Case
authors.forename: Alan
authors.initials: A
authors.lastname: Korman
authors.forename: Xiao-Yong
authors.initials: XY
authors.lastname: Li
authors.forename: Amelia
authors.initials: A
authors.lastname: Black
authors.forename: David J
authors.initials: DJ
authors.lastname: King
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D024861:Genetic Enhancement / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014157:Transcription Factors / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D014157:Transcription Factors