pNiFty3-T-Lucia

pNiFty3-T-Lucia plasmid is composed of three key elements: the mouse interferon beta minimal promoter, five NFAT transcription factor binding sites and a secreted luciferase (Lucia) reporter gene.

pNiFty3-T-Lucia plasmid is selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones

Specifications

Transcription factor binding sites: NFAT (4x)
Minimal Promoter: mouse IFNβ promoter
Selection: Zeocin™
Reporter Gene: Lucia luciferase

These products are covered by a Limited Use License (See Terms and Conditions).

Contents

• 20 µg of lyophilized DNA
• 1 ml of Zeocin™ (100 mg/ml)

Product is shipped at room temperature.

Description

Minimal promoter

The proximal promoters are shorter than 500    bp and contain transcription  factor binding sites. Upon stimulation   in  293 cells, their expression  level remains undetectable. With the    addition of repeated TFBS, the  proximal promoters become inducible by    the appropriate stimulus and  drive the expression of the reporter  gene.

IFN-β promoter: the mouse IFN-β minimal  promoter comprises several positive regulatory domains that bind  different cooperating transcription factors such as NF-kB, IRF3 and IRF7  [1].

Transcription factor binding sites (TFBS)

NFAT  binding site: Nuclear factor of activated T-cell (NFAT) is a family of  transcription factors expressed in T cells, but also in other classes of  immune and non-immune cells [2]. NFAT is activated by stimulation of  receptors coupled to calcium mobilization, such as the PRRs Dectin-1 and  Mincle [3,4]. Calcium mobilization induces the calmodulin-dependent  phosphatase calcineurin leading to NFAT activation. NFAT binds to a 9 bp  element, with the consensus sequence (A/T)GGAAA(A/N)(A/T/C)N.

Reporter Gene

Lucia luciferase reporter gene: Lucia luciferase is a secreted luciferase with strong bioluminescent activity
The     Lucia reporter gene is ideal for promoter activity and gene     expression  studies. Lucia is encoded by a synthetic gene derived by     genetic  engineering of copepod luciferase genes. Lucia luciferase     activity can  be detected and quantified directly in the culture medium     of  transfected cells using InvivoGen's QUANTI-Luc™ detection reagent.

1.  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.
2. Rao A. et al., 1997. Transcription factors of the NFAT family: regulation and function. Annu Rev Immunol. 15:707-47.
3.  Goodridge HS. et al., 2007. Dectin-1 stimulation by Candida albicans  yeast or zymosan triggers NFAT activation in macrophages and dendritic  cells. J Immunol. 178(5):3107-15.
4. Yamasaki S. et al., 2009. C-type  lectin Mincle is an activating receptor for pathogenic fungus,  Malassezia. PNAS. 106(6):1897-902.

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