Identification and Use of Zinc Finger Transcription Factors That Increase Production of Recombinant Proteins in Yeast and Mammalian Cells

2008 ◽  
Vol 21 (3) ◽  
pp. 664-670 ◽  
Author(s):  
Kyung-Soon Park ◽  
Wongi Seol ◽  
Hyo-Young Yang ◽  
Seong-Il Lee ◽  
Sung Keun Kim ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Zinc Finger ◽  
Recombinant Proteins ◽  
Mammalian Cells

scholarly journals COMET: A toolkit for composing customizable genetic programs in mammalian cells

2019 ◽  
Author(s):  
Patrick S. Donahue ◽  
Joseph W. Draut ◽  
Joseph J. Muldoon ◽  
Hailey I. Edelstein ◽  
Neda Bagheri ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mathematical Model ◽  
Transcription Factors ◽  
Zinc Finger ◽  
Mammalian Cells ◽  
Single Layer ◽  
Performance Characteristics ◽  
Boolean Logic ◽  
Design And Construction

ABSTRACTEngineering mammalian cells to carry out sophisticated and customizable genetic programs requires a toolkit of multiple orthogonal and well-characterized transcription factors (TFs). To address this need, we developed the COmposable Mammalian Elements of Transcription (COMET)—an ensemble of TFs and promoters that enable the design and tuning of gene expression to an extent not previously possible. COMET currently comprises 44 activating and 12 inhibitory zinc-finger TFs and 83 cognate promoters, combined in a framework that readily accommodates new parts. This system can tune gene expression over three orders of magnitude, provides chemically inducible control of TF activity, and enables single-layer Boolean logic. We also develop a mathematical model that provides mechanistic insights into COMET performance characteristics. Altogether, COMET enables the design and construction of customizable genetic programs in mammalian cells.

scholarly journals A protocol for rapid degradation of endogenous transcription factors in mammalian cells and identification of direct regulatory targets

2021 ◽  
Vol 2 (2) ◽  
pp. 100530
Author(s):  
Hillary M. Layden ◽  
Nicholas A. Eleuteri ◽  
Scott W. Hiebert ◽  
Kristy R. Stengel
Keyword(s):  
Transcription Factors ◽  
Mammalian Cells ◽  
Rapid Degradation

scholarly journals Expression Analyses of Soybean VOZ Transcription Factors and the Role of GmVOZ1G in Drought and Salt Stress Tolerance

2020 ◽  
Vol 21 (6) ◽  
pp. 2177 ◽  
Author(s):  
Bo Li ◽  
Jia-Cheng Zheng ◽  
Ting-Ting Wang ◽  
Dong-Hong Min ◽  
Wen-Liang Wei ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Salt Stress ◽  
Abiotic Stress ◽  
Stress Tolerance ◽  
Zinc Finger ◽  
Hairy Roots ◽  
Yeast Cells ◽  
Salt Stress Tolerance ◽  
Drought And Salt Stress

Vascular plant one-zinc-finger (VOZ) transcription factor, a plant specific one-zinc-finger-type transcriptional activator, is involved in regulating numerous biological processes such as floral induction and development, defense against pathogens, and response to multiple types of abiotic stress. Six VOZ transcription factor-encoding genes (GmVOZs) have been reported to exist in the soybean (Glycine max) genome. In spite of this, little information is currently available regarding GmVOZs. In this study, GmVOZs were cloned and characterized. GmVOZ genes encode proteins possessing transcriptional activation activity in yeast cells. GmVOZ1E, GmVOZ2B, and GmVOZ2D gene products were widely dispersed in the cytosol, while GmVOZ1G was primarily located in the nucleus. GmVOZs displayed a differential expression profile under dehydration, salt, and salicylic acid (SA) stress conditions. Among them, GmVOZ1G showed a significantly induced expression in response to all stress treatments. Overexpression of GmVOZ1G in soybean hairy roots resulted in a greater tolerance to drought and salt stress. In contrast, RNA interference (RNAi) soybean hairy roots suppressing GmVOZ1G were more sensitive to both of these stresses. Under drought treatment, soybean composite plants with an overexpression of hairy roots had higher relative water content (RWC). In response to drought and salt stress, lower malondialdehyde (MDA) accumulation and higher peroxidase (POD) and superoxide dismutase (SOD) activities were observed in soybean composite seedlings with an overexpression of hairy roots. The opposite results for each physiological parameter were obtained in RNAi lines. In conclusion, GmVOZ1G positively regulates drought and salt stress tolerance in soybean hairy roots. Our results will be valuable for the functional characterization of soybean VOZ transcription factors under abiotic stress.

scholarly journals DNA-binding specificity of GATA family transcription factors.

1993 ◽  
Vol 13 (7) ◽  
pp. 3999-4010 ◽  
Author(s):  
M Merika ◽  
S H Orkin
Keyword(s):  
Transcription Factors ◽  
Dna Binding ◽  
Mammalian Cells ◽  
Chimeric Protein ◽  
Binding Specificity ◽  
Mobility Shift ◽  
Binding Domains ◽  
Dna Binding Specificity ◽  
Mobility Shift Assay ◽  
Gata Family

GATA-binding proteins constitute a family of transcription factors that recognize a target site conforming to the consensus WGATAR (W = A or T and R = A or G). Here we have used the method of polymerase chain reaction-mediated random site selection to assess in an unbiased manner the DNA-binding specificity of GATA proteins. Contrary to our expectations, we show that GATA proteins bind a variety of motifs that deviate from the previously assigned consensus. Many of the nonconsensus sequences bind protein with high affinity, equivalent to that of conventional GATA motifs. By using the selected sequences as probes in the electrophoretic mobility shift assay, we demonstrate overlapping, but distinct, sequence preferences for GATA family members, specified by their respective DNA-binding domains. Furthermore, we provide additional evidence for interaction of amino and carboxy fingers of GATA-1 in defining its binding site. By performing cotransfection experiments, we also show that transactivation parallels DNA binding. A chimeric protein containing the finger domain of areA and the activation domains of GATA-1 is capable of activating transcription in mammalian cells through GATA motifs. Our findings suggest a mechanism by which GATA proteins might selectively regulate gene expression in cells in which they are coexpressed.

Effects of different zinc finger transcription factors on genomic targets

2006 ◽  
Vol 339 (1) ◽  
pp. 263-270 ◽  
Author(s):  
Leon W. Neuteboom ◽  
Beatrice I. Lindhout ◽  
Ingrid L. Saman ◽  
Paul J.J. Hooykaas ◽  
Bert J. van der Zaal
Keyword(s):  
Transcription Factors ◽  
Zinc Finger
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