CHOP Responsive Luciferase Reporter Mia-Paca2 Stable Cell Line is derived from human pancreatic cancer, and stably express firefly luciferase reporter gene under the control of the Unfolded Protein Response, ER stress response element. This cell line is an ideal cellular model for monitoring the activation of Unfolded Protein Response, ER stress Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.
CHOP, also known as GADD153, is a multifunctional transcription factor of the Unfolded Protein Response or endoplasmic reticulum stress (ER stress). During UPR, the activation of PERK-ATF4 pathway upregulates CHOP protein level, although IRE1 and ATF6 pathways can also regulate CHOP expression. Transcriptionally active CHOP increases target genes, such as Ero1 and GADD34, and mediates ER stress-induced apoptosis. CHOP also plays an important role in protein synthesis, intracellular calcium regulation, and oxidation during ER stress.
Signosis has developed CHOP luciferase reporter stable cell line by transducing cells with baculovirus containing both CHOP luciferase reporter and hygromycin expression cassette. The hygromycin resistant clones were subsequently screened for thapsigargin-induced luciferase activity. The cell line can be used as a reporter system for monitoring the activity of CHOP triggered by stimuli treatment, such as thapsigargin, tunicamycin, gene overexpression and gene knockdown. The cells contain no viral particles and require handling at biosafety level 1 protocol.
Principle behind TF luciferase reporter. TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression. The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites. There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways. Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression. Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.
Analysis of SL-0025 CHOP reporter activity in response to thapsigargin treatment. The Mia-Paca2 cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with or without 300nM thapsigargin in DMEM and 10% FBS for 16 hours. Mia-Paca2-CHOP Luciferase Reporter Cell Line exhibits dose-dependent increase in luciferase activity when compared to untreated cells.