NFAT Luciferase Reporter Lentivirus
The NFAT Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the NFAT response element located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (hygromycin, puromycin, or geneticin) for the selection of stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Figure 1. Schematic of the lenti-vector used to generate the NFAT luciferase reporter lentivirus (hygromycin).
Figure 1. Schematic of the lenti-vector used to generate the NFAT luciferase reporter lentivirus (puromycin).
Figure 1. Schematic of the lenti-vector used to generate the NFAT luciferase reporter lentivirus with geneticin (NeoR/KanR) selection.
Name | Ordering Information |
Jurkat cells | ATCC #TIB-152 |
Anti-CD3 agonist antibody | BPS Bioscience #71274 |
Thaw Medium 2 | BPS Bioscience #60184 |
Polybrene | Millipore, #TR-1003-G |
96-well tissue culture treated white clear-bottom assay plate | Corning, #3610 |
One-Step luciferase assay system | BPS Bioscience #60690 |
Luminometer |
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
NFAT (Nuclear factor of activated T-cells) is a family of transcription factors that has an important function in immune responses, for example by inducing the expression of various cytokines (such as IL-2-3-4 and TNF-alpha) in T cells. Members of the NFAT family have been found in many tissue types including heart, skeletal muscle and brain cells. Through their role in the immune system NFATs are involved in inflammation and these transcription factors are considered promising therapeutic targets for a variety of diseases.
NFAT is regulated by Ca2+ and the Ca2+/calmodulin-dependent serine phosphatase calcineurin. NFAT proteins are phosphorylated and reside in the cytoplasm in resting cells; upon stimulation, they are dephosphorylated by calcineurin, translocate to the nucleus, and induce gene expression.
- Clipstone NA, Crabtree GR. Nature. 1992 Jun 25;357(6380):695-7.
- Lyakh, L., et al. Mol Cell Biol. 1997 May;17(5):2475-84.