pGreenFire1 (pGF1) is a versatile HIV-based lentivector that co-expresses destabilized copGFP and Firefly Luciferase enabling the detection of both GFP signals as well as Luciferase for quantitative transcription activation reporter assays.

Features and Benefits of pGreenFire1 (pGF1) Lentivectors

● pGreenFire1 is a dual reporter vector and so has the advantage of being able to determine whether reporter activation has taken place by assessing both GFP and firefly luciferase levels.

● Firefly luciferase provides a familiar, well-established and quantitative assessment of transcription activation, while the GFP reporter enables non-destructive live cell imaging.

● Proviral reporters are integrated into normal chromatin and therefore provide a more accurate model for the study of transactivation and epigenetic effects.

 ● pGreenFire1 lentiviral reporters allow researchers to quickly and easily establish stable reporter cell lines using either their own transcriptional response elements or any of SBI’s pre-made, pre-validated reporters.

● Stable reporter cell lines eliminate the need to transfect reporter plasmids for each experiment and so reduces the well-to-well variability often seen with plasmid reporter transfection.

● No need to have a second reporter (eg Renilla luciferase or beta-galactosidase) to normalize transfection levels.

● Minimal CMV (mCMV) basal promoter has extremely low activity in the absence of transcription response elements.

Sample Data using the pGreenFire1-NF-κB Transcription Reporter

 ● Monitoring NF-κB transactivation using GFP epifluorescence

293TN cells were transduced with the packaged pGF1-NFκB reporter virus. Two days later, the cells were split into 96 well plates and then treated with the indicated amount of TNF-α. Levels of NFκB activation was assessed 18 hours post-induction using epifluorescence microscopy.               

 ● Quantitative analysis of NF-κB transactivation using Firefly Luciferase

293TN cells were transduced with either the packaged pGF1-NFκB or pGF1-mCMV (negative control) reporter virus. Two days later, the cells were split into 96 well plates and then treated with the indicated amount of TNF-α. Levels of NFκB activation was quantitated 18 hours post-induction using a standard firefly luciferase assay (Promega).               

Kit Components

Each kit contains 10 μg of endotoxin-free plasmid DNA in TE buffer or >2 x 10⁶ IFUs pre-packaged lentivirus.

Each kit comes with a product certificate of analysis.
 

User Manual

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Ordering Information

 ● pGreenFire1 Lentivectors

●  PA-1 product version refers to plasmid (10 μg)

●  VA-1 product version refers to virus (>2 x 10⁶ IFUs)

● pGreenFire1 Pre-made Transcription Reporter Constructs

●  PA-1 product version refers to plasmid (10 μg)

All pre-made reporter plasmids come with:

1. pGreenFire1-CMV positive control plasmid (PA, 10 μg)

2. pGreenFire1 negative control plasmid (PA, 10 μg)

3. pGreenFire1 + TRE plasmid (PA, 10 μg)

●  VA-1 product version refers to virus (>2 x 10⁶ IFUs)

Control viruses available (see top table) for discounted price; please inquire

To obtain more information contact SBI at 888-266-5066 (toll-free in North America) or 650-968-2200, or e-mail to orders@systembio.com

Details of the Lentiviral Delivery System:

• Simple packaging into pseudoviral particles with SBI’s pPACK Lentivector Packaging System (see: http://www.systembio.com/pPACK.htm)

• Deliver pGreenFire1 lenticonstructs into a wide range of cell lines including non-dividing or difficult-to-transfect cell lines with self-inactivating and high-titer enabled HIV-based lentivector.

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