Using a secreted and robust Gaussia Luciferase (GLuc) as the reporter, GeneCopoeia GLuc-ON™ promoter clones are designed for promoter analysis by detecting the real-time activities of over 20,000 human and 18,000 mouse promoters using live cell assays.
Each transfection-ready promoter clone contains a 1.2-1.5 kb insert, corresponding to the 5'-flanking promoter sequence located approximately 1.5 kb upstream and up to 200 bp downstream of the transcription start site (TSS) of a specific human gene. This insert is placed upstream of the GLuc reporter gene. Since the putative cis-acting enhancer elements are expected to exist in the cloned promoter region, the promoter luciferase activity observed during the reporter assay closely resembles the actual promoter regulation of these genes within human cells.
GLuc-ON promoter clones can be ordered as pre-designed or custom-built in one of our robust vector systems. Currently we offer both single-reporter and dual-reporter vector systems. The single-reporter system uses GLuc, mCherry, or GFP as the promoter reporter. The dual-reporter system uses GLuc as the promoter reporter and SEAP (secreted Alkaline Phosphatase) as the internal control for signal normalization.
Figure 1. How GLuc-ON promoter clones work.
Advantages |
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Live cell assays
- Naturally secreted GLuc reporter
- No lysis of the cells is necessary
- Save samples, reduce variations, and simplify experiments for applications such as pulse chase analysis,etc.
Real-time study
- Data is generated quickly
- Closely resembles real-time activities
Dual secreted reporter system
- Secreted GLuc and SEAP
- Enables transfection-normalization for accurate across-sample comparison
High-throughput compatible
- Group or pathway study compatible
- High sample number compatible
High sensitivity
- GLuc is 1000-fold more sensitive than firefly or Renilla luciferase
Convenience
- All promoter clones are transfection-ready
Gaussia luciferase
GLuc-ON promoter clones use a modified GLuc (mGLuc) as the reporter gene, which generates a highly stable signal and overcomes the quick signal decay commonly observed with humanized wild type GLuc (wtGLuc).
Figure 2. Signal stability of mGLuc (blue) and wtGLuc (red). Left: assay buffer with a stabilizer; Right: regular assay buffer (Secrete-Pair™ dual luminescence assay kit)
Dual-reporter system
Dual-reporter vectors are available for the GLuc-ON promoter clones. The secondary reporter, secreted Alkaline Phosphatase (SEAP), serves as an internal control. The dual-reporter system enables transfection normalization for accurate cross-sample comparison.
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Figure 3. Normalized promoter activities in H1B1B and HEK293T cells. Dual-reporter promoter clones or controls were transfected into two cell lines in duplicates. Samples were analyzed 24 hrs (HEK293T) and 48 hrs (H1B1B) after transfection. NEG (containing non-promoter sequence) and EMPTY (no promoter in the vector) are negative controls. |
Vector choices |
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| Vector | Reporter gene | Tracking gene | Selection marker | Vector type |
| pEZX-PG04** | Gaussia luciferase (GLuc) | Secreted alkaline phosphatase (SEAP) | Puromycin | Non-viral |
| pEZX-PG02** | Gaussia luciferase (GLuc) | N/A* | Puromycin | Non-viral |
| pEZX-PF02 | eGFP | N/A* | Puromycin | Non-viral |
| pEZX-PM02 | mCherry | N/A* | Puromycin | Non-viral |
| pEZX-LvPG04** | Gaussia luciferase (GLuc) | Secreted alkaline phosphatase (SEAP) | Puromycin | HIV Lentiviral |
| pEZX-LvPG02** | Gaussia luciferase (GLuc) | N/A* | Puromycin | HIV Lentiviral |
| pEZX-LvPF02 | eGFP | N/A* | Puromycin | HIV Lentiviral |
| pEZX-LvPM02 | mCherry | N/A* | Puromycin | HIV Lentiviral |
* A separate vector is available for SEAP expression.
** These Gaussia luciferase cloning vectors are available for purchase.
Control clones and SEAP expression clone
Tear Away License
Pre-designed promoter clonesSearch over 20,000 human and 18,000 mouse pre-designed promoter reporter clones in your choice of vector.
Promoter lentivirusGeneCopoeia offers lentivirus as a custom service.
Custom promoter clone services
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Want to compare various length or regions of your promoter? You can custom-order them.
Please complete our Custom Promoter Clone Inquiry and Quotation form,
email inquiry@genecopoeia.com or call: 866-360-9531 or 301-762-0888
| Download warranty |
| GLuc-ON™ Promoter Reporter Clones Cancellation, QA, License, and Warranty |
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- MethylAffinity™ Methylated DNA Enrichment Kit
Promoter reporter clone publications
- Zhang, H. et al. (2014) The critical role of myeloid-derived suppressor cells and FXR activation in immune-mediated liver injury. Journal of Autoimmunity, doi: 10.1016/j.jaut.2014.02.010. [MPRM11909-PG04]
- Rubie, C. et al. (2014) Chemokine receptor CCR6 expression is regulated by miR-518a-5p in colorectal cancer cells. Journal of translational medicine, 2014 Feb 21;12:48. doi: 10.1186/1479-5876-12-48.. [Luc CCR6 vector, Luc No CCR6 vector]
- Klimosch, SN. et al (2013) Functional TLR5 genetic variants affect human colorectal cancer survival. Cancer Research, 73(24); 7232–42.. [Promoter Reporter ( MYD88 and ACAA1)].
- Shen, Q. et al (2013) Adipocyte reporter assays: Application for identification of anti‐inflammatory and antioxidant properties of mangosteen xanthones. Molecular Nutrition & Food Research, DOI: 10.1002/mnfr.201300181. [Human Fabp4 promoter vector with eCFP reporter].
- Celardo, I. et al. (2013) Caspase-1 is a novel target of p63 in tumor suppression. Cell Death and Disease (2013) 4, e645; doi:10.1038/cddis.2013.175. [caspase-1 promoter plasmid in pEZX-PG02 vector].
- Zheng, H. et al. (2013) Glycogen synthase kinase-3 beta regulates Snail and β-catenin expression during Fas-induced epithelial–mesenchymal transition in gastrointestinal cancer. European Journal of Cancer, Available online 9 April 2013. [CDH1 promoter construct].
- Mills, L.D. et al. (2013) Loss of the Transcription Factor GLI1 Identifies a Signaling Network in the Tumor Microenvironment Mediating KRAS-Induced. Journal of Biological Chemistry, March 12, 2013, doi: 10.1074/jbc.M112.438846. [mouse SHH promoter clone].
- Schank, JR. et al (2013) Tacr1 Gene Variation and Neurokinin 1 Receptor Expression Is Associated with Antagonist Efficacy in Genetically Selected Alcohol-Preferring Rats. Biological Psychiatry Journal, PII: S0006-3223(13)00043-7, doi:10.1016/j.biopsych.2012.12.027. [custom Tacr1 promoter luciferase construct].
- Petrella, B. et al. (2012) Interleukin-1 beta and transforming growth factor-beta 3 cooperate to activate matrix metalloproteinase expression and invasiveness in A549 lung adenocarcinoma … Cancer Letters, Volume 325, Issue 2, 28 December 2012, Pages 220–226. [MMP10 promoter clone].