PMID- 23685433
OWN - NLM
STAT- MEDLINE
DCOM- 20130920
LR  - 20211021
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 41
IP  - 13
DP  - 2013 Jul
TI  - Synthetic mammalian trigger-controlled bipartite transcription factors.
PG  - e134
LID - 10.1093/nar/gkt405 [doi]
AB  - Synthetic biology has significantly advanced the design of synthetic control 
      devices, gene circuits and networks that can reprogram mammalian cells in a 
      trigger-inducible manner. Prokaryotic helix-turn-helix motifs have become the 
      standard resource to design synthetic mammalian transcription factors that tune 
      chimeric promoters in a small molecule-responsive manner. We have identified a 
      family of Actinomycetes transcriptional repressor proteins showing a tandem 
      TetR-family signature and have used a synthetic biology-inspired approach to 
      reveal the potential control dynamics of these bi-partite regulators. Daisy-chain 
      assembly of well-characterized prokaryotic repressor proteins such as TetR, ScbR, 
      TtgR or VanR and fusion to either the Herpes simplex transactivation domain VP16 
      or the Krueppel-associated box domain (KRAB) of the human kox-1 gene resulted in 
      synthetic bi- and even tri-partite mammalian transcription factors that could 
      reversibly program their individual chimeric or hybrid promoters for 
      trigger-adjustable transgene expression using tetracycline (TET), 
      gamma-butyrolactones, phloretin and vanillic acid. Detailed characterization of the 
      bi-partite ScbR-TetR-VP16 (ST-TA) transcription factor revealed independent 
      control of TET- and gamma-butyrolactone-responsive promoters at high and double-pole 
      double-throw (DPDT) relay switch qualities at low intracellular concentrations. 
      Similar to electromagnetically operated mechanical DPDT relay switches that 
      control two electric circuits by a fully isolated low-power signal, TET programs 
      ST-TA to progressively switch from TetR-specific promoter-driven expression of 
      transgene one to ScbR-specific promoter-driven transcription of transgene two 
      while ST-TA flips back to exclusive transgene 1 expression in the absence of the 
      trigger antibiotic. We suggest that natural repressors and activators with tandem 
      TetR-family signatures may also provide independent as well as DPDT-mediated 
      control of two sets of transgenes in bacteria, and that their synthetic 
      transcription-factor analogs may enable the design of compact therapeutic gene 
      circuits for gene and cell-based therapies.
FAU - Folcher, Marc
AU  - Folcher M
AD  - Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 
      CH-4058 Basel, Switzerland.
FAU - Xie, Mingqi
AU  - Xie M
FAU - Spinnler, Andrea
AU  - Spinnler A
FAU - Fussenegger, Martin
AU  - Fussenegger M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130517
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Bacterial Proteins)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Herpes Simplex Virus Protein Vmw65)
RN  - 0 (Kruppel-Like Transcription Factors)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (ScbR protein, Streptomyces coelicolor)
RN  - 0 (Trans-Activators)
RN  - 0 (ZNF10 protein, human)
RN  - 0 (tetracycline resistance-encoding transposon repressor protein)
SB  - IM
MH  - Actinobacteria/genetics
MH  - Amino Acid Motifs
MH  - Bacterial Proteins/chemistry/genetics
MH  - Cells, Cultured
MH  - DNA-Binding Proteins/chemistry/genetics
MH  - *Gene Expression Regulation
MH  - HEK293 Cells
MH  - Herpes Simplex Virus Protein Vmw65/genetics
MH  - Humans
MH  - Kruppel-Like Transcription Factors/genetics
MH  - Promoter Regions, Genetic
MH  - Protein Engineering/methods
MH  - Recombinant Fusion Proteins/chemistry
MH  - Repressor Proteins/*chemistry/classification/genetics/metabolism
MH  - Synthetic Biology/methods
MH  - Trans-Activators/*chemistry/genetics/metabolism
MH  - Transgenes
PMC - PMC3711444
EDAT- 2013/05/21 06:00
MHDA- 2013/09/21 06:00
PMCR- 2013/05/17
CRDT- 2013/05/21 06:00
PHST- 2013/05/21 06:00 [entrez]
PHST- 2013/05/21 06:00 [pubmed]
PHST- 2013/09/21 06:00 [medline]
PHST- 2013/05/17 00:00 [pmc-release]
AID - gkt405 [pii]
AID - 10.1093/nar/gkt405 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2013 Jul;41(13):e134. doi: 10.1093/nar/gkt405. Epub 2013 May 
      17.