pNiFty, a collection of inducible SEAP or luciferase reporter plasmids
InvivoGen has designed the pNiFty plasmid collection to monitor pattern recognition receptor (PRR) and cytokine signaling through the inducible expression of reporter genes. You may choose between the inducible reporter gene encoding either for SEAP (secreted alkaline phosphatase), or luciferases including Lucia®, Firefly, or Renilla.
Two families of pNiFty plasmids are available:
— pNiFty2-N family: NF-κB-inducible reporter plasmids
— pNiFty3-I family:IRF-inducible reporter plasmids
The promoter region of the pNiFty plasmids is composed of the following key elements:
- a proximal promoter
- repeated transcription factor binding sites (TFBS)
pNiFty2 family:
The pNiFty2-N plasmids feature an engineered ELAM (endothelial cell-leukocyte adhesion molecule) promoter combined with NF-κB TFBS to enhance the NF-κB-mediated transcription [1]. These pNiFty2-N plasmids are selectable with Zeocin® in both E. coli and mammalian cells and can be used to generate stable clones. They are also available with either Blasticidin or Puromycin resistance.
pNiFty3 family:
The pNiFty3-I plasmids feature the mouse IFN-β minimal promoter combined with several ISRE (interferon responsive elements) TFBS to enhance the IRF-mediated transcription [2]. These pNiFty3 plasmids are selectable with Blasticidin in both E. coli and mammalian cells and can be used to generate stable clones. They are also available with either Zeocin® or Puromycin resistance.
► pNifTy2-N and pNiFty3-I families allow you to generate your own Dual reporter cell lines for the simultaneous monitoring of NF-κB and IRF activation, using the combination of reporter and antibiotic resistance genes that suits you best.
Learn more about creating your own Dual reporter cell lines.
Advantages of pNiFty plasmids:
- Large choice of reporter genes encoding for SEAP or luciferases, including Lucia®, Firefly, or Renilla
- Available with different antibiotic resistances: Zeocin®, Blasticidin, and Puromycin
- Selectable in both E.coli and mammalian cells
- Suitable for the generation of stable clones
- Combined use of pNiFty2-N and pNiFty3-I for simultaneous monitoring of NF-κB and IRF signaling
For your convenience, InvivoGen also provides the reporter detection reagents QUANTI-Blue™, QUANTI-Luc™ 4 Gaussia/Lucia, and QUANTI-Luc™ 4 Renilla to measure SEAP, Lucia®, and Renilla activities, respectively.
Note: Lucia® is a trademark of InvivoGen registered with the United States Patent and Trademark Office.
References:
1. Schindler U, Baichwal VR, 1994. Three NF-kappa B binding sites in the human E-selectin gene are required for maximal tumor necrosis factor alpha-induced expression. Mol Cell Biol. 14(9):5820-31.
2. Vodjdani G. et al., 1988. Structure and characterization of a murine chromosomal fragment containing the interferon beta gene. J Mol Biol. 204(2):221-31.
