Description:

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.

Principle:

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.

 

Data:

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.

Signosis 是一家专业的生物分析供应商，致力于基于微孔板分析技术的生命科学研究产品的开发、生产和销售。Signosis 成立于2007年，坐落于世界研发中心--美国硅谷的中心地带。公司专注于细胞因子、转录因子、miRNA及疾病标志物等重要调控因子和标志物的研究，基于自有专利技术，开发完成400多种先进而独特的用于发现、筛选和分析的生命科学研究产品。Signosis 产品销售遍布北美、欧洲、亚洲及中东等主要国家和地区，客户涵盖美国国立卫生研究院、哈佛大学、耶鲁大学等著名研究机构以及众多制药公司