Monitor signal transduction in real time with our re-engineered pGreenFire 2.0 Lentivectors

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

*Please note that these vectors only function properly when transduced. Transfection keeps the constitutive RSV promoter intact, leading to nonspecific expression of the reporter genes. 相比传统的四环素病毒调控系统，SBI（system biosciences）最新推出的SparQ™ Cumate   调控系统无疑将为很多利用病毒载体做研究的实验者带来福音。这一系统通过不断的病毒转导，实   现严谨调控，强有力的反馈和可检测的表达，其主要用于诱导调控基因和microRNA的表达。这一   系统通过结合在cumate操纵子上的高亲和能力的CymR阻遏物进行调控，通过加入一种可以结合   在CymR上的无毒的小分子化合物Cumate，来解除抑制。这一系统可以反复使用，精确的开关机   制，从而实现严谨的调控，用作时空表达的研究。                       图一：无渗漏调控系统原理图   相比传统的诱导系统，Cumate 具有更低的背景和强劲的表达，如图二所示，相比其他的系统，SparQ比其他系统表达强32倍，且是零渗漏。  同时SBI还可以根据您的需要定制Cumate系统！