NFAT Pathway

1. About NFAT Pathway:

NFAT (Nuclear Factor of Activated T-cells) proteins are transcription factors that play essential roles in T-cell activation, development, and function, as well as in other cell types. They are regulated by their phosphorylation status, which affects their localization and activity. NFAT signaling pathway is activated by an increase in intracellular calcium levels, which dephosphorylates NFAT proteins. Dephosphorylated NFAT translocates from the cytoplasm to the nucleus, binds to specific DNA sequence, the transcriptional response element (TRE) in the promoter, leading to the activation of its target genes [Ref1].

2. Lentivirus Detects NFAT Pathway:

GenTarget developed a set of reporting lentivirus to monitor NFAT signaling pathway activity. Those reporting lentivirus has a luminescent report (Luciferase) or a fluorescent report (GFP), under a minimal CMV promoter (mCMV) that embedded with optimized tandem repeats of NFAT pathway-specific transcriptional response element (TRE), sequence motif (5′- GGAGGAAAAAC-TGTTTCATACAGAAGGCGT-TGGAAACTC   3′) [Ref 2].

NFAT pathway is activated primarily by signals that increase intracellular calcium levels. Upon activated (dephosphorylates), the NFAT proteins bind to the TRE in report’s promoter region, leading to increased report expression, the Luc or GFP.

3. Structure of the Reporting Lentivirus:

The pathway specific reporting Lentivirus use either firefly-luciferase or GFP signal changes to detect the pathway’s activity.   Those reporting lentivirus also contains a constitutively expressed Puromycin antibiotic selection marker, or a GFP fluorescent selection marker, which makes it easier to select the stably transduced signal reporting cells, via Puromycin killing or GFP sorting respectively. See lentivector core scheme below.

The premade, ready-to-use reporter lentivirus provides a much easier tool to monitor the activity of the signaling pathways in virtually any mammalian cell type. It also allows to generate your own reporting cell line in your desired cell type for study or screening of pathway specific gene-knockdown, over-expression, or chemical / drug/protein treatment in the cell based assay.

Click the Product Manual (pdf) for all available products for this pathway reporter.