Description(s):

ATF6 Responsive Luciferase Reporter CHO-K1 Stable Cell Line is derived from Chinese Hamster Ovary , and stably express firefly luciferase reporter gene under the control of ATF6 response element.  This cell line is an ideal cellular model for monitoring the activation of Unfolded Protein, ER  stress Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.

Principle

ATF6 plays a key role in the Unfolded Protein Response, as ATF6 acts synergistically with other ER stress sensors, PERK and IRE1, to mitigate ER stress.  Upon activation, ATF6 translocates to the nucleus and binds to cis-acting ER stress element that is present in promoters of ER chaperones.

Signosis has developed ATF6 luciferase reporter stable cell line by transfecting cells with plasmids containing ATF6 luciferase reporter and hygromycin expression cassette.  The hygromycin resistant clones were subsequently screened for thapsigargin or tunicamycin-induced luciferase activity.   The cell line can be used as a reporter system for monitoring the activity of ATF6 triggered by stimuli treatment, gene overexpression and gene knockdown.

 

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-0024 ATF6 reporter activity in response to ER stress. The CHO-K1 cells were seeded on a 96-well plate for overnight with DMEM/F12 including 10% FBS. The cells then were treated with indicated concentrations of tunicamycin (TM) or thapsigargin (TG) in DMEM/F12 and 10% FBS for 16 hours.  CHO-K1-ATF6 Luciferase Reporter Cell Line exhibits stress-dependent increase in luciferase activity when compared to control cells. Data was read on a luminometer with a sensitivity of 3x10^-21 moles luciferase.

 

 

Literature

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Citations

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Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.   NRF2/ARE Luciferase Reporter NIH3T3 Stable Cell Line (SL-0047)   Thiamine and Benfotiamine Protect Neuroblastoma Cells against Paraquat and β-Amyloid Toxicity by a Coenzyme-Independent Mechanism. Sambon, Margaux, et al. Heliyon, vol. 5, no. 5, 2019, doi:10.1016/j.heliyon.2019.e01710.   NFkB Luciferase Reporter HEK293 Stable Cell Line (SL-0012)   Vitale LA, He LZ, Thomas LJ, et al. Development of CDX-527: a bispecific antibody combining PD-1 blockade and CD27 costimulation for cancer immunotherapy. Cancer Immunology, Immunotherapy : CII. 2020 May. DOI: 10.1007/s00262-020-02610-y.   Luciferase Substrate (LUC15/100)   Direct Antioxidant Properties of Methotrexate: Inhibition of Malondialdehyde-Acetaldehyde-Protein Adduct Formation and Superoxide Scavenging. Matthew C. Zimmermana, Dahn L. Clemensb, c, Michael J. Duryeeb, Cleofes Sarmientoa, Andrew Chioub, Carlos D. 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