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 insertion of multiple repeats of a response element upstream of a minimal promoter (minP).
To construct pathway reporters, the pGL4.26[luc2/minP/Hygro], pGL4.27[luc2P/minP/Hygro] and pGL4.28[luc2CP/minP/Hygro] Vectors have minimal promoter (minP) vectors with three varieties of engineered firefly luciferase genes. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P and luc2CP 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, and the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. These vectors also have a hygromycin resistance selectable marker for use either in transient transfection experiments or selection of a stable cell line.
- Transcription regulation.
- Virus-cell interactions.
- Compound screening.
- Post-translational modifications.
- GPCR signaling.
- Cell signaling.
About the pGL4 Reporter Vectors
The pGL4 Vectors offer increased reporter gene expression with codon optimization of synthetic genes for mammalian expression and reduced background and risk of expression artifacts with removal of cryptic DNA regulatory elements and transcription factor binding sites. Firefly luciferase has options that offer improved temporal response with the destabilized Rapid Response™ luciferase genes.