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  • Gaussia Luciferase

    Gaussia Luciferase (GLuc) is a recently discovered, naturally secreted protein by the copepod, Gaussia princeps (1). At 19kD, it is the smallest known luciferase and its catalytic properties make it one of the “brightest” known (2). Additionally, multiple disulfide bonds contribute to the protein stability at elevated temperatures. These properties make it an ideal reporter gene, either as a standalone expression monitor or as a fusion partner with other proteins (3). The first report of in vivo imaging using Gluc demonstrated sensitivity that is several orders of magnitude higher than the first generation luciferases, Firefly Luciferase (Fluc) and Renilla (4). In addition to mammalian cells, the secretion signal of GLuc is functional in all eukaryotic cells, including algae and nematodes. Scientists at NEB have used Gaussia luciferase to develop siRNA potency and miRNA expression assays (5,6). The GLuc reporter is increasingly being used to monitor the cellular secretory pathway and ER stress, to assay for viral infectivity and to study chemokine receptor activation (3,7-10).

    Activity of Gaussia (GLuc) and Cypridina (CLuc) Luciferases Expressed in CHO Cells.

    Activity was determined from the conditional media or cell lysate.

    Extreme Sensitivity of the Gaussia Luciferase Reporter.

    HeLa cells were transfected with 1 µg of pCMV-GLuc or pCMV-Renilla, and activity was measured from the cell supernatant (S) or cell lysate (L). Note that although Gaussia Luciferase is a secreted reporter, there is sufficient measurable activity inside the cell. Renilla is only expressed intracellularly.

    References

    1. Balou, B., Szent-Gyorgyi, C. and Finley, G (2000) Abstract from 11th Int. Symp. on Biolumin. & Chemilumin. Asilomar, CA. 
    2. Goerke, A., Loening, A., Gambhir, S. and Swartz, J. (2008) Metabolic Engineering, 10, 187–200. PMID: 18555198
    3. Hulleman JD et al.(2011) Mol. Biol. Cell December 15, 2011 vol. 22 no. 244765-4775. PMID: 22031286 
    4. Tannous, B.A. et al (2005) Mol. Ther. 11, 435-443. PMID: 15727940
    5. Morlighem, J.E., Petit, C and Tzertzinis, G. (2007) Biotechniques, 42, 599-606. PMID: 17515198
    6. Shea C. and Tzertzinis G. unpublished observations (2007). 
    7. Badr, et al. (2007) PloS ONE, 2, e571. PMID: 17593970
    8. Hewett, J.W., et al. (2007) Proc. Natl. Acad. Sci, USA, 104, 7271-7276. PMID: 17428918
    9. Capul, A. and de la Torre, J.C. (2008) Virology 382, 107-114. PMID: 18929379
    10. Luker, K., Gupta, M. and Luker G. (2009) Biotechniques, 47, 625-632. PMID: 19594447

    Featured Products

    Advantages

    • Naturally Secreted – Amenable to live cell assays
    • Sensitivity – Brightest luciferases available; enables single cell applications
    • Stability – Samples can be stored for several days with no loss of activity
    • Easy-to-use – Cell lysis not necessary
    • Non-destructive – Living cells can be used in downstream assays
    • Flexible – Activity can also be measured in cell lysates

    Applications

    • Transfection optimization studies
    • Promoter/enhancer assays
    • High throughput assays
    • Multiplex assays
    • Multiple assays with other reporters
    • Secretory pathway reporter assays
    • Signal transduction
    • siRNA potency screening
    • Time course studies
    • Single cell assays, including stem cells and primary cells
    • Live cell assays
    • Assays in difficult to transfect cells

    NEB vs. Other Commercially Available Reporter Systems

    Features Gaussia Cypridina Renilla Firefly Metridia
    Transcriptional
    Reporter
    Extreme Sensitivity      
    Secreted    
    Stable      
    Does not require and is
    not affected by ATP
     

    Comparison of the features of commercially available reporter systems highlights the advantages of using Gaussia or Cypridina Luciferase from NEB.

    Reporter System Selection Chart

    NEB offer a variety of products that utilize secreted Gaussia Luciferase (GLuc) and Cypridina Luciferase (CLuc) in reporter assays.