pGreenFire 2.0 NFAT reporter plasmid (pGF2-NFAT-rFluc-T2A-GFP-mPGK-Puro)

SBI has upgraded our popular pGreenFire signaling pathway reporter lentivectors with a design that leads to more reliable generation of stable cell lines. We’ve also swapped in the red firefly luciferase reporter (rFLuc), which opens up the possibility of performing a dual-spectral luciferase assay and delivers greater sensitivity for in vivo applications than conventional luciferase.

With the pGreenFire 2.0 NFAT Reporter Lentivector & Virus (pGF2-NFAT-rFluc-T2A-GFP-mPGK-Puro) the core reporter functionality is similar to the original pGreenFire lentivector—NFAT transcriptional response elements (TREs) are placed upstream of a minimal CMV promoter (mCMV) which together drive co-expression of rFLuc and GFP in response to NFAT activity. The result is the ability to quantitatively measure NFAT activity using both fluorescence and luciferase activity.

What makes our next-gen pGreenFire 2.0 vectors even better than other TRE reporter vectors is the smart design, which adds in a constitutive selection cassette for stable cell line generation while minimizing interference with the upstream TRE. By using a weak/moderate mPGK promoter to drive the antibiotic selection marker (puromycin resistance) and carefully arranging the conditional reporter genes, the selection marker is reliably expressed without compromising conditional expression of rFLuc and GFP.

As with our original pGreenFire1 vectors, all pGreenFire 2.0 lentivectors leverage our reliable lentivector technology and save you time with pre-built signal transduction pathway reporters that come as ready-to-transduce pre-packaged lentivirus and plasmid that can be transfected into the lentivirus producing system of your choice*.

• Sort responsive cells with GFP
• Measure activity with red firefly luciferase
• Leverage SBI’s highly-regarded lentivectors
• Create stable signaling pathway reporter cell lines
• Introduce reporters into difficult-to-transfect cell types, including primary and non-dividing mammalian cell lines

Supporting Data

See our pGreenFire 2.0 transcriptional response element reporters in action

 

Figure 1. The pGreenFire 2.0 NFAT Reporter efficiently and quantitatively reports on NFAT activity in Jurkat cells. Relative luciferase activity (A) and GFP activity (B) both increase in response to PMA and ionomycin, which together induce NFAT activity. Induction is specific to the NFAT transcription response elements as the pGreenFire 2.0 mCMV Negative Control displays very little luciferase activity regardless of the presence of inducer.