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 Overview Advantages Applications
NEB offers several products that utilize secreted Gaussia Luciferase (GLuc) and Cypridina Luciferase (CLuc) in reporter assays. These systems offer extreme sensitivity and can be used alone or together as a dual system.
Gaussia Luciferase (GLuc) is a 20 kDa luciferase from the marine cocepod Gaussia princeps. This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that produces light (480 nm), and has considerable advantages over other luminescent reporter genes. It is normally secreted from the cells and its secretion signal also functions very efficiently in mammalian cells. GLuc offers the advantage of a greatly increased bioluminescent signal relative to the commonly used firefly (Fluc) and Renilla luciferases (RLuc). GLuc was determined to emit light with a specific activity of 4.2 x 1024 photons/s/mol, the highest reported activity for any characterized luciferase (1).
Cypridina Luciferase (CLuc) is isolated from the marine ostracod Cypridina noctiluca and is efficiently secreted from mammalian cells (2,3). CLuc does not react with coelenterazine, a common substrate of marine luciferases. It uses a specific luciferin, which is not a substrate for Gaussia Luciferase. This allows the detection of both of GLuc and CLuc expressed from the same cells.
- Verhaegan, M.and Christopoulos, T.K. (2002) Anal. Chem. 74, 4378-4385.
- Wu, C. Chie, Suzuki-Ogoh, C., Ohmlya, Y. (2007) Biotech. 42, 290-292.
- Nakajima, et al. (2004) Biosci. Biotechnol. Biochem. 68, 565–570.
Activity of Gaussia (GLuc) and Cypridina
(CLuc) luciferases expressed in CHO cells.
Activity was determined from the conditional
media or cell lysate. |
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
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Extreme Sensitivity of the Gaussia Luciferase
Reporter. HeLa cells were transfected with 1 µg
of pCMV-GLuc or pCMV-Renilla, and light
emission
was measured for the 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.
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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
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Comparison of the features of commercially available reporter systems highlights the advantages of using Gaussia or Cypridina Luciferase from NEB
| Features |
Gaussia |
Cypridina |
Renilla |
Firefly |
Metridia |
Transcriptional
Reporter |
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| Extreme Sensitivity |
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√ |
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| Secreted |
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√ |
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| Stable |
√ |
√ |
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Does not require and
is
not affected by ATP |
√ |
√ |
√ |
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Technical Reference Quick Links |
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