Enhanced protein production by engineered zinc finger proteins.
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.
Reik
Andreas
A
Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Richmond, CA 94804, USA.
Zhou
Yuanyue
Y
Collingwood
Trevor N
TN
Warfe
Lyndon
L
Bartsevich
Victor
V
Kong
Yanhong
Y
Henning
Karla A
KA
Fallentine
Barrett K
BK
Zhang
Lei
L
Zhong
Xiaohong
X
Jouvenot
Yann
Y
Jamieson
Andrew C
AC
Rebar
Edward J
EJ
Case
Casey C
CC
Korman
Alan
A
Li
Xiao-Yong
XY
Black
Amelia
A
King
David J
DJ
Gregory
Philip D
PD
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*
D011994:Recombinant Proteins; D014157:Transcription Factors