Regulatory

Part:BBa_K4421021

Designed by: Shuyang Zhu   Group: iGEM22_NMU_China   (2022-06-03)
Revision as of 03:32, 9 June 2022 by Juejue (Talk | contribs) (supplementary information)


NFAT-RE promotor

This part is a CAR inducible promoter containing six NFAT-RE sites in a minimal IL-2 promoter to generate CAR response elements, which was placed upstream of the transcription factor Gal4-KRAB.In our engineered NK-92 cells, NFAT was coupled with KRAB activation after CAR stimulation.

Usage and Biology

This part can serve as a downstream promotor of CAR recognition in synthetic immunology, especially in constructing CAR-classis cells with unique circus.Together with synnotch,we can really do a lot.

supplementary information

It consists of six NFAT-RE sites. NFAT-RE can bind with NFAT factors, thus activating expression of the downstream genes, which is original from the IL-2 promotor. The regulation of IL-2 gene was revealed comprehensively during last century. Several factors are required for induction of the IL-2 gene after stimulation, including NFAT1, NF-KB, AP-1, and octamer-binding proteins(shapiro et al.,1994). It was previously reported that synthetic NFAT promoter could be successfully applied in CAR-T cells (Chinnasamy D et al.,2012) .However,in NK-92 cells, a previous study showed that reporter expression and fold induction from the 4xNFAT promoter was very modest (kulemzin, S.V. et al, 2019). So we designed the NFAT-RE with 6xNFAT and the expression test with GFP showed that it did work in our project. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None