Creating a cell line with an indicator of a functional signaling pathway is useful for deciphering the components in a signaling pathway. These tools are made by inserting multiple repeats of a response element upstream of a minimal promoter (minP). We have designed vectors that report the activity of a variety of pathways using the optimized luc2 firefly luciferase gene in the pGL4 backbone. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P and RapidResponse™ genes are luc2 genes appended with degradation sequences to influence the cellular half-life of firefly luciferase. Proteins encoded by the RapidResponse™ genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity.
These response element vectors also have a hygromycin resistance selectable marker for use either in transient transfection experiments or for selection of a stable cell line. Some of these response element vectors also are available stable cell lines (GloResponse™ Cell Lines) to speed your research. For investigating the cytokine signaling pathway, we have vectors available include that respond to interferon-? (INF-?) with the pGL4.45[luc2P/ISRE/Hygro] Vector, interleukin 6 (IL-6) with the pGL4.47[luc2P/SIE/Hygro] Vector, transforming growth factor ? (TGF-?) with the pGL4.48[luc2P/SBE/Hygro] Vector and interleukin 3 (IL-3) with the pGL4.52[luc2P/STAT5RE/Hygro] Vector.
- Transcription regulation.
- Virus-cell interactions.
- Compound screening.
- Post-translational modifications.
- GPCR signaling.
- Cell signaling.
Activator/Pathway Transcription Factor Binding Site Vector Name Example Data INF-? STAT1:STAT2 Interferon Stimulated Response Element (ISRE) pGL4.45[luc2P/ISRE/Hygro] TGF-? SMAD3:SMAD4 SMAD Binding Element (SBE) pGL4.48[luc2P/SBE/Hygro] IL6 STAT3:STAT3 sis-Inducible Element (SIE) pGL4.47[luc2P/SIE/Hygro] IL3 STAT5:STAT5 STAT5 Response Element pGL4.52[luc2P/STAT5 RE/Hygro]
pGL4.47[luc2P/SIE/Hygro] Vector Protocol
PDF (195 KB)
pGL4.48[luc2P/SBE/Hygro] Vector Protocol
PDF (192 KB)
pGL4.52[luc2P/STAT5RE/Hygro] Vector Protocol
PDF (208 KB)
pGL4.45[luc2P/ISRE/Hygro] Vector Protocol
PDF (215 KB)
Certificate of AnalysisLookup Certificate of Analysis
-30C TO -10C
For Research Use Only. Not for Use in Diagnostic Procedures.