Part:BBa_K4803118

Introduction

This biobrick is created by combining BBa_K4803007(STAT3 responsive element), BBa_K4803010(mini promoter), BBa_K3190204(ZsGreen).

Usage and Biology

This Part consists of a mini promoter and ZsGreen downstream of the STAT3 responsive element, which promotes ZsGreen expression under conditions of AP-1 activation. IL-6 induces activation of STAT3 [1]. This Part, when introduced into human cells, fluoresces in response to IL-6 concentration, making it useful for visualizing IL-6 concentration.

UTokyo2023 project needed to confirm the function of the Modular Extracellular Sensor Architecture (MESA) system [2], which detects IL-6, and whether the concentration of IL-6 is sufficiently altered by inhibitors secreted by the Secretion system. Therefore, we thought we could quantify the concentration of IL-6 by monitoring STAT3.

Experimental results

Figure 1: Mode of fluorescence intensity for each concentration of IL-6 in iU62 (reporter cell for IL-6) at 24h after IL-6 addition. Error bar indicates standard error

Figure 2: Graph of fluorescence intensity per concentration of IL-6 in iU62 (reporter cell for IL-6) at 24 h after addition of IL-6. Error bar indicates standard error.

Blue → 0 pg/ml　Red → 100 pg/ml

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Wang, Y., Boxel-dezaire, A. V., Cheon, H., & Stark, G. R. (2013). STAT3 Activation in Response to IL-6 Is Prolonged by the Binding of IL-6 Receptor to EGF Receptor. BIOLOGICAL SCIENCES, 110(42), 16975–16980. https://doi.org/https://doi.org/10.1073/pnas.1315862110

[2] Daringer, N. M., Dudek, R. M., Schwarz, K. A., & Leonard, J. N. (2014). Modular extracellular sensor architecture for engineering mammalian cell-based devices. ACS synthetic biology, 3(12), 892-902. https://doi.org/10.1021/sb400128g

[3] Xu, J., Lin, H., Wu, G., Zhu, M., & Li, M. (2021). IL-6/STAT3 is a promising therapeutic target for hepatocellular carcinoma. Frontiers in oncology, 11, 760971. https://doi.org/10.3389/fonc.2021.760971

[4]Wei, W., Tweardy, D. J., Zhang, M., Zhang, X., Landua, J., Petrovic, I., Bu, W., Roarty, K., Hilsenbeck, S. G., Rosen, J. M., & Lewis, M. T. (2014). STAT3 signaling is activated preferentially in Tumor‐Initiating cells in Claudin‐Low models of human breast cancer. Stem Cells (Dayton, Ohio), 32(10), 2571-2582. https://doi.org/10.1002/stem.1752