Part:BBa_K4803117

Introduction

This biobrick is created by combining BBa_K4803008(AP-1 responsive element), BBa_K4803010(mini promoter), BBa_K3190204(ZsGreen).

Usage and Biology

This Part encodes the mini promoter and ZsGreen downstream of the AP-1 responsive element, which promotes ZsGreen expression under conditions of AP-1 activation. IL-1β induces activation of AP-1 and NF-κB [1]. This Part, when transfected into human cell lines, fluoresces in response to IL-1β concentrations, making it useful for visualizing IL-1β concentration.

In UTokyo2023 project, we needed to confirm the function of the MESA system to detect IL-1β and whether the concentration of IL-1β is sufficiently altered by inhibitors secreted by the Secretion unit. Therefore, we thought it would be possible to quantify the concentration of IL-1β by monitoring AP-1.

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]

Reference

[1] Wolf, J. S., Chen, Z., Dong, G., Sunwoo, J. B., Bancroft, C. C., Capo, D. E., Yeh, N. T., Mukaida, N., & Waes, C. V. (2001). IL (Interleukin)-1α Promotes Nuclear Factor-ΚB and AP-1-Induced IL-8 Expression, Cell Survival, and Proliferation in Head and Neck Squamous Cell Carcinomas. Clin Cancer Res, 7(6), 1812–1820. https://aacrjournals.org/clincancerres/article/7/6/1812/288814/IL-Interleukin-1-Promotes-Nuclear-Factor-B-and-AP

[2] Jutz, S., Leitner, J., Schmetterer, K., Doel-Perez, I., Majdic, O., Grabmeier-Pfistershammer, K., Paster, W., Huppa, J. B., & Steinberger, P. (2016). Assessment of costimulation and coinhibition in a triple parameter T cell reporter line: Simultaneous measurement of NF-κB, NFAT and AP-1. Journal of Immunological Methods, 430, 10-20. https://doi.org/10.1016/j.jim.2016.01.007

[3] Invivogen. (n.d.). IL-1β Reporter HEK 293 Cells. Products, InvivoGen. https://www.invivogen.com/hek-blue-il6