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Nuclear Receptor Reporter Stable Cell Lines
Nuclear Hormone Receptors (NHRs) represent a fascinating class of ligand-binding transcription factors (TFs) crucial in regulating gene expression. With over 350 NHRs cataloged in the Protein Data Bank, their intricate role in cellular processes is underscored by the modulation of activities through specific ligand interactions. While conventional DNA regulatory element-luciferase reporter cell lines have been instrumental in studying these receptors, their limitations arise from a lack of specificity due to shared binding sequences among family members. Recognizing this challenge, Signosis has innovatively devised ligand-binding domain (LBD)-driven GAL4 reporter stable cell lines, a method poised to offer heightened sensitivity, specificity, and reduced toxicity. This transformative approach opens avenues for studying NHRs in a more nuanced and precise manner, particularly in drug compound screening for agonist or antagonist properties.
Principle
Nuclear hormone receptors (NHRs) constitute a group of ligand-binding transcription factors (TFs). More than 350 NHRs are available in the Protein Data Bank. Like other TFs, they can regulate gene expression by binding to specific DNA regulatory elements, but their activities are modulated by the corresponding ligands.
Although DNA regulatory element-luciferase reporter cell lines can be used to study endogenous or exogenous receptors and analyze the receptor signaling pathway in a native biological context, the NHR DNA element-luciferase reporter cell lines lack specificity and uniqueness due to the similarity in binding sequences among their entire family members.
Signosis has developed ligand-binding domain (LBD)-driven GAL4 reporter stable cell lines by delivering two vectors, one expressing 8 copies of GAL4 upstream activator sequences (UAS) in front of the luciferase reporter gene, and another expressing Gal4 DNA Binding Domain (DBD) bound to a ligand binding domain (LBD) of your choice. Upon addition of a corresponding ligand, the DBD-LBD fusion protein is activated, binding to the GAL promoter binding site that will drive forward the expression of luciferase. With this method, our GAL4-based cell line assays will have minimal cross-reactivity with other nuclear receptors, leading to high sensitivity and specificity, while also having low toxicity of the chimeric receptors to the cells when overexpressed. These cell lines can be used to screen drug compounds for agonist or antagonist.
Stable clones were selected with both hygromycin and G418, and a functional assay was subsequently conducted, with clones exhibiting the highest induction fold chosen.
Benefits
Highly Specific
The responsiveness of this stable cell line is driven by the specific ligand-binding domain with low/no cross-reactivity with other members of the nuclear receptor.
Routine Mycoplasma Testing
All cell lines tested negative for mycoplasma.
Less Toxic
The chimeric receptors display less toxic effects on the cells.
We also offer our own Firefly Luciferase Substrate which can generate a strong light signal that is comparable to our competitor's, but at a more cost-effective price.
Highly Sensitive
More than 50 fold induction in response to the corresponding stimuli.
Ultra Sensitive
NR LBD-DRIVEN GAL 4
Cell Lines
The Ultra Sensitive LBD-driven GAL4 reporter cell lines has been optimized with an enhanced promoter upstream of the luciferase gene to maximize signal output in response to extremely low concentrations of inducers. These cell line not only yields the enhanced basal signal suitable for any instrument detection, but also produces the higher induction to monitor the specific pathways in an ultra sensitive manner.
Ultra Sensitive GAL4
NR LBD-Driven GAL4
Cell Lines
Signosis has developed ready-to-use Nuclear Receptor Ligand Binding Domain driven GAL4 reporter cell lines.
These cell lines are a reliable, rapid, and sensitive method for analyzing the intracellular status of NRs upon exposure to drug candidates or other stimuli. These functional assays allow for measurement of receptor activation or inhibition by compounds and can be used to determine compound potency and selectivity.
GAL4-UAS-Luc
Cell Lines
Signosis has developed the GAL4-UAS-Luc Reporter Stable Cell Lines as a reliable, rapid, and sensitive method for analyzing the expression of your target gene of interest upon exposure to related ligands or other stimuli. These cell lines are transfected only with the GAL4-UAS-LUC vector, and are ready to be transfected with your customized GAL4 vector containing your NR of interest.
GAL4-UAS-LUC
Research Application
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Studying the effects of ligand binding on GAL4-mediated transcriptional activity
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Identifying ligands that activate or inhibit the target receptors by monitoring GAL4-driven luciferase expression
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Screening for potential therapeutic compounds that modulate the target receptors by measuring changes in luciferase activity
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Investigating the mechanisms underlying the regulation of the target receptors by analyzing the GAL4-mediated transcriptional response to various stimuli
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Comparing the activity and selectivity of different compounds on the target receptors by using the GAL4 reporter assay
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Studying the interaction between the target receptors and other signaling pathways or co-regulators by analyzing the changes in GAL4-mediated transcriptional activity upon co-expression of different genes or treatments.
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Studying the effects of ligand binding on PPAR-gamma-mediated transcriptional activity
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Identifying compounds that activate or inhibit PPAR-gamma activity by monitoring GAL4-driven luciferase expression
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Investigating the role of PPAR-gamma in adipogenesis and glucose homeostasis by analyzing the GAL4-mediated transcriptional response to various stimuli
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Screening for potential therapeutic compounds that target PPAR-gamma for the treatment of diabetes and other metabolic disorders
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Studying the interaction between PPAR-gamma and other signaling pathways or co-regulators by analyzing the changes in GAL4-mediated transcriptional activity upon co-expression of different genes or treatments.
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Studying the effects of ligand binding on PPAR-alpha or PPAR-delta-mediated transcriptional activity
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Identifying compounds that activate or inhibit PPAR-alpha or PPAR-delta activity by monitoring GAL4-driven luciferase expression
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Investigating the roles of PPAR-alpha and PPAR-delta in lipid metabolism and energy homeostasis by analyzing the GAL4-mediated transcriptional response to various stimuli
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Screening for potential therapeutic compounds that target PPAR-alpha or PPAR-delta for the treatment of metabolic disorders, including dyslipidemia and obesity
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Studying the interaction between PPAR-alpha or PPAR-delta and other signaling pathways or co-regulators by analyzing the changes in GAL4-mediated transcriptional activity upon co-expression of different genes or treatments.
