Part:BBa_K4421021
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 a unique circus. Together with synnotch, we can do a lot.
Supplementary Information
It consists of six NFAT-RE sites. NFAT-RE can bind with NFAT factors, thus activating the expression of the downstream genes, which is original from the IL-2 promoter. The regulation of the IL-2 gene was revealed comprehensively during the 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 were very modest (Kulemin, 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Reference
Shapiro et al.(1994).c-Rel Regulation of IL-2 Gene Expressionism May Be Mediated Through Activation of AP-1.Journal of Experimental Medicine, 184(5):1663-1669.https://doi.org/10.1084/jem.184.5.1663
Chinnasamy D et al.(2012).Local delivery of interleukin-12 using T cells targeting VEGF receptor-2 eradicates multiple vascularized tumors in mice.Clinical Cancer Research,2012;18:1672–83.https://doi.org/10.1158/1078-0432.CCR-11-3050
Kulemzin et al.(2019).Design and analysis of stably integrated reporters for inducible transgene expression in human T cells and CAR NK- cell lines.BMC Medical Genomics,12(Suppl 2):44.https://doi.org/10.1186/s12920-019-0489-4
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