Introduction

GeneCopoeia’s GLuc-ON™ EGR1 transcriptional response element (TRE) lentiviral clone enables analysis of the effects on genes regulated by early growth response protein 1 (EGR1), such as transforming growth factor-beta-1 (TGFB1) and fibronectin (FN1). EGR1 is a transcription factor that responds to mitogenic stimulation. EGR1 regulates many processes, such as neuron activity, wound healing, apoptosis, and angiogenesis, and is known to both suppress and promote tumor growth.

The GLuc-ON™ PKC EGR1 lentiviral clone contains tandem repeats of EGR1 upstream of a minimal promoter and a secreted Gaussia luciferase reporter gene, allowing for robust and sensitive analysis without cell lysis. The EGR1 response elements are engineered to provide strong activation with low background (Figure 1). The GLuc-ON™ EGR1 TRE lentiviral clone is provided as transfection-ready DNA. A negative (non-activating) control lentiviral clone is also available for purchase.

User Manual

Validation Data

Figure 1. Activation of the GLuc-ON™ EGR1 TRE by phorbol 12-myristate 13-acetate (PMA). HEK293a cells were transduced with negative control lentiviral particles or the GLuc-ON™ EGR1 TRE lentiviral particles. Transduced cells were incubated for 48 hours, followed by 6 days of incubation in the presence of puromycin. Puromycin-resistant cells were then treated with PMA for 18 hours. A Gaussia Luciferase assay was performed, and transcriptional response activity values are expressed as the fold activation of luminescence.

Introduction

GeneCopoeia’s GLuc-ON™ PKC transcriptional response element (TRE) lentiviral clone enables analysis of the effects on genes regulated by activator protein 1 (AP1). AP1 is a transcription factor complex comprised of the c-Fos and c-Jun family proteins,  and is necessary for signaling through the Protein kinase C (PKC) pathway.

The GLuc-ON™ PKC TRE lentiviral clone contains tandem repeats of PKC upstream of a minimal promoter and secreted Gaussia luciferase reporter gene, which allows robust and sensitive analysis without lysing the cells.  The PKC response elements are engineered to provide strong activation with low background (Figure 1). The GLuc-ON™ PKC TRE lentiviral clone is provided as transfection-ready DNA. A negative (non-activating) control lentiviral clone is also available for purchase.

User Manual

Validation Data

Figure 1. Activation of the GLuc-ON™ PKC TRE by phorbol 12-myristate 13-acetate (PMA). HEK293a cells were transduced with negative control lentiviral particles or the GLuc-ON™ PKC TRE lentiviral particles. Transduced cells were incubated for 48 hours, followed by 6 days of incubation in the presence of puromycin. Puromycin-resistant cells were then treated with PMA for 18 hours. A Gaussia Luciferase assay was performed, and transcriptional response activity values are expressed as the fold activation of luminescence.  

Introduction

GeneCopoeia provides the GLuc-ON™ TCF/LEF transcriptional response element (TRE) lentiviral clone for analysis of the effects on Wnt signaling pathways mediated by TCF/LEF transcription factors, which recruit β-catenin to activate the promoters of several genes critical for function of the Wnt pathway. Wnt signaling plays fundamental roles in embryonic development, and its misregulation has been implicated in diseases such as cancer and type 2 diabetes.

The GLuc-ON™ TCF/LEF TRE lentiviral clone contains tandem repeats of TCF/LEF upstream of a minimal promoter and secreted Gaussia luciferase reporter gene, which allows robust and sensitive analysis without lysing the cells.  The TCF/LEF response elements are engineered to provide strong activation with low background (Figure 1). The GLuc-ON™ TCF/LEF TRE lentiviral clone is provided as transfection-ready DNA. A negative (non-activating) control lentiviral clone is also available for purchase.

User Manual

Validation Data

Figure 1. Activation of the GLuc-ON™ Wnt TRE by lithium chloride. HEK293a cells were transduced with negative control lentiviral particles or the GLuc-ON™ TCF/LEF TRE lentiviral particles. Transduced cells were incubated for 48 hours, followed by 6 days of incubation in the presence of puromycin. Puromycin-resistant cells were then treated with lithium chloride for 18 hours. A Gaussia  Luciferase assay was performed, and transcriptional response activity values are expressed as luminescence fold activation.  

Introduction

GeneCopoeia’s GLuc-ON™ NFAT transcriptional response element (TRE) lentiviral clone enables analysis of calcium signaling pathways mediated by the nuclear factor of activated T cells (NFAT) family of transcription factors. NFATs, in combination with AP-1, regulate several genes, such as cytokines, in response to levels of intracellular calcium, and are important functional determinants of the immune system.

The GLuc-ON™ NFAT TRE lentiviral clone contains tandem repeats of NFAT upstream of a minimal promoter and secreted Gaussia luciferase reporter gene, which allows robust and sensitive analysis without lysing the cells.  The NFAT response elements are engineered to provide strong activation with low background (Figure 1). The GLuc-ON™ NFAT TRE lentiviral clone is provided as transfection-ready DNA. A negative (non-activating) control lentiviral clone is also available for purchase.

User Manual

Validation Data

Figure 1. Activation of the GLuc-ON™ NFAT TRE by PMA + ionomycin. HEK293a cells were transduced with negative control lentiviral particles or the GLuc-ON™ NFAT TRE lentiviral particles. Transduced cells were incubated for 48 hours, followed by 6 days of incubation in the presence of puromycin. Puromycin-resistant cells were then treated with PMA and ionomycin for 18 hours. A Gaussia Luciferase assay was performed, and transcriptional response activity values are expressed as the fold activation of luminescence.  

Introduction

GeneCopoeia’s GLuc-ON™ CREB transcriptional response element (TRE) lentiviral clone enables analysis of the effects on the cAMP and PKA signaling pathways through the cAMP response element (CRE). CRE, which is found in the promoters of many genes, including somatotatin, vegetative growth factor (VGF), and the protooncogene c-fos, becomes bound by the transcription factor CRE binding protein (CREB) in response to elevated cAMP levels.

The GLuc-ON™ CREB TRE lentiviral clone contains tandem repeats of CRE upstream of a minimal promoter and secreted Gaussia luciferase reporter gene, which allows robust and sensitive analysis without lysing the cells.  The CREB response elements are engineered to provide strong activation with low background (Figure 1). The GLuc-ON™ CREB TRE lentiviral clone is provided as transfection-ready DNA. A negative (non-activating) control lentiviral clone is also available for purchase.

User Manual

Validation Data

Figure 1. Activation of the GLuc-ON™ CREB TRE by forskolin. HEK293a cells were transduced with  negative control lentiviral partices or the GLuc-ON™ CREB TRE lentiviral particles. Transduced cells were incubated for 48 hours, followed by 6 days of incubation in the presence of puromycin. Puromycin-resistant cells were then treated with forskolin for 18 hours. A Gaussia  Luciferase assay was performed, and transcriptional response activity values are expressed as luminescence fold activation.