Description(s):

Amino acid deprivation-AARE Luciferase Reporter MEF Stable Cell Line is derived from murine embryonic fibroblast, and stably express firefly luciferase reporter gene under the control of AARE (Amino acid response element).  This cell line is an ideal cellular model for monitoring Amino acid response Signaling Pathway triggered by amino acid starvation or stimuli treatment.

Principle

Besides the central role of amino acids as building blocks of proteins they are also involved in other cellular functions like transcription, mRNA maturation, translation, mRNA turnover, autophagy and metabolism. Lack of amino acids (deprivation) will trigger the amino acid response (AAR) pathway largely through signal transduction and mediated through altered transcriptional activity of specific genes, including eIF2α, Asns, Atf4 and Chop.

Signosis has developed a luciferase reporter stable cell line under control of AARE (Amino acid response element) that can be easily used to monitor activation of Amino acid response Signaling Pathway.

 

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 Amino acid resp-AARE Luciferase Reporter MEF Stable Cell Line in response to stimuli. The cells were seeded on a 96-well plate overnight with DMEM including 10% FBS. The cells then were treated with or without 300 nM TG (Thapsigargin) in DMEM and 0.1% FBS for 16 hours. Then cells were lysed and RLU was read. Data was read on a luminometer with a sensitivity of 3x10^-21 moles luciferase.

Literature

View user manual

 

Citations

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Genes Cancer. 2016 Nov; 7(11-12): 368–382., doi:  10.18632/genesandcancer.128   Diallyl trisulfide inhibits proliferation, invasion and angiogenesis of glioma cells by inactivating Wnt/β-catenin signaling. Qingxia Tao, Cuiying Wu, Ruxiang Xu, Lijun Niu, Jiazhen Qin, Ning Liu, Peng Zhang, Chong Wang. Cell and Tissue Research, 2017, doi:10.1007/s00441-017-2678-9.   BCN057 induces intestinal stem cell repair and mitigates radiation-induced intestinal injury. Payel Bhanja, Andrew Norris, Pooja Gupta-Saraf, Andrew Hoover and Subhrajit Saha. Stem Cell Research & Therapy, vol. 9, no. 1, Feb. 2018, doi:10.1186/s13287-017-0763-3.   (Nrf2/ARE) luciferase reporter cell line HEK293 (SL-0042)   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. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018   Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad1,*, Geetha Venkateswaran1,*, Li Hao1, Maria E. Garcia1, Jenny Yoon1, Jaskiran Sidhu1 and Sujata Persad1.Genes & Cancer. January 30, 2017 . https://doi.org/10.18632/genesandcancer.128    HIF Luciferase Reporter Hela Stable Cell Line (SL-0023)   Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.   NFkB Luciferase Reporter HepG2 Stable Cell Line (SL-0017)   Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.   CHOP Luciferase Reporter Mia_Paca2 Stable Cell Line (SL-0025)   A new ent-norabietant diterpenoid from roots of Euphorbia lathyris L. Yun, Young Sook, et al.  Tetrahedron Letters, 2018, doi:10.1016/j.tetlet.2018.06.017.   NFAT Luciferase Reporter Jurkat Stable Cell Line (SL-0032)   Diversification of Bw4 Specificity and Recognition of a Nonclassical MHC Class I Molecule Implicated in Maternal–Fetal Tolerance by Killer Cell Ig-like Receptors of the Rhesus Macaque. Banerjee, Priyankana, et al. The Journal of Immunology, 2018, doi:10.4049/jimmunol.1800494.   P329G-CAR-J: a Novel Jurkat-NFAT-Based CAR-T Reporter System Recognizing the P329G Fc Mutation. Darowski, Diana, et al. Protein Engineering, Design and Selection, 2019, doi:10.1093/protein/gzz027.   Dendritic cells dictate responses to PD-L1 blockade cancer immunotherapy. Mayoux, Maud, et al.  Science Translational Medicine Vol. 12, Issue 534, 2020. DOI: 10.1126/scitranslmed.aav7431    NRF2/ARE Luciferase Reporter HepG2 Stable Cell Line (SL-0046)   Innovative drinking water treatment techniques reduce the disinfection-induced oxidative stress and genotoxic activity. Lundqvist, Johan, et al. Water Research, vol. 155, 2019, pp. 182–192., doi:10.1016/j.watres.2019.02.052.   Stat3 Luciferase Reporter Hela Stable Cell Line (SL-0003)   Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.   An engineered ligand trap inhibits leukemia inhibitory factor as pancreatic cancer treatment strategy. Hunter, S.A., McIntosh, B.J., Shi, Y. et al. Commun Biol 4, 452 (2021). https://doi.org/10.1038/s42003-021-01928-2   Stat1 Luciferase Reporter Hela Stable Cell Line (SL-0004)   Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. 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. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018   FXR Luciferase Reporter HepG2 Stable Cell Line (SL-0055) Targeting the Liver‐Brain Axis with Hop‐Derived Flavonoids Improves Lipid Metabolism and Cognitive Performance in Mice. Paraiso, I.L., Revel, J.S., Choi, J., Miranda, C.L., Lak, P., Kioussi, C., Bobe, G., Gombart, A.F., Raber, J., Maier, C.S. and Stevens, J.F. Mol. Nutr. Food Res. 2020, 2000341. https://doi.org/10.1002/mnfr.202000341   Estrogen Receptor Luciferase Reporter T47D Stable Cell Line (SL-0002)   Glass-bottled drinking water: a time capsule to study the historic presence of hazardous chemicals using effect-based methods. Lundqvist, J., Persson, K.M. & Oskarsson, A. Environ Sci Eur 33, 34 (2021). https://doi.org/10.1186/s12302-021-00476-0