Regulatory

Part:BBa_K4160006

Designed by: Femi Hesen, Wouter Langers, Floor van Boxtel   Group: iGEM22_TU-Eindhoven   (2022-10-09)
Revision as of 14:16, 11 October 2022 by Fvboxtel (Talk | contribs)


STAT promoter + CMVmin

This part encodes for the STAT promoter, which can bind STAT proteins. Upon binding of STAT proteins, STAT-induced genes will be expressed.1 In this part, the STAT promoter is accompanied by a cytomegalovirus (CMV) sequence. This sequence is a commonly used enhancer element that allows for the high-level production of recombinant proteins in mammalian cells.2


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 53
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 53
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 53
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 53
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

The STAT promoter is a DNA sequence that regulates the transcription of downstream STAT-induced genes.1 The STAT promoter induces gene transcription upon binding of the transcription factor STAT (BBa_K4160005). These STAT proteins bind to DNA recognition motifs, named gamma-activated sites (GAS), that are located in the STAT promoter sequence. This binding initiates the transcription of downstream genes.3


GEMS system

TU-Eindhoven 2022 used this part as a transcription factor for the Generalized Extracellular Molecule Sensor (GEMS) system. Upon dimerization of the GEMS receptor, IL-6RB (BBa_K4160002) domains are activated and induce downstream signaling of the JAK/STAT pathway, which results in STAT-induced gene expression.


Characterization

Characterization of expression of STAT-induced proteins in the GEMS system can be found on the BBa_K4160008 page.



References

  1. Lee DS, Grandis JR, Johnson DE. STAT3 as a Major Contributor to Chemoresistance. Target Cell Surviv Pathways to Enhanc Response to Chemother. Published online 2019:145-167. doi:10.1016/B978-0-12-813753-6.00007-X
  2. Wang W, Jia YL, Li YC, et al. RETRACTED ARTICLE: Impact of different promoters, promoter mutation, and an enhancer on recombinant protein expression in CHO cells. Sci Reports 2017 71. 2017;7(1):1-10. doi:10.1038/s41598-017-10966-y
  3. Aaronson DS, Horvath CM. A road map for those who don’t know JAK-STAT. Science (80- ). 2002;296(5573):1653-1655. doi:10.1126/SCIENCE.1071545/ASSET/F0B466F3-364A-4A06-96A1-AF7833D916CE/ASSETS/GRAPHIC/SE2020533001.JPEG




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