Reporter Systems

New England Biolabs offers a selection of products for the study and manipulation of gene expression. These products include a group of validated and highly efficient transfection reagents for the most commonly used cell lines, as well as many primary endothelial and epithelial cells.

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.

Detection of Low Numbers of Cells Expressing GLuc and CLuc

The indicated number of cells expressing GLuc or CLuc was diluted with untransfected CHO cells and cultured in a 24-well plate format for 1 day. A 20 µl sample from each well was assayed for each luciferase.

Featured Products

Reporter Systems includes these subcategories:

Gaussia Luciferase Information
Cypridina Luciferase Information

    Publications related to Reporter Systems:

  1. Zhu, W. et al. (2011). A reporter system for assaying influenza virus RNP functionality based on secreted Gaussia luciferase activity Virology J. . 10.1186/1743-422X-8-29.
  2. Watanabe, T., Enomoto, T., Takahashi, M., Honma, S., Honma, K. and Ohmiya, Y. (2010). Multichannel perfusion culture bioluminescence reporter system for longterm detection in living cells Anal. Biochem. .
  3. Kaihara, A., Sunami, A., Kurokawa, J., Furukawa, T. (2009). A genetically encoded bioluminescent indicator for the sodium channel activity in living cells J. Am. Chem. Soc.. 1, 4188-4189.
  4. Kaihara, A., Sunami, A., Kurokawa, J. and Furukawa, T. (2009). A genetically encoded bioluminescent indicator for the sodium channel activity in living cells J. Am. Chem. Soc. . 1, 4188-4189.
  5. Ding, L., Paszkowski-Rogacz, M., Nitzsche, A., Slabicki, M.M., Heninger, A.K., de Vries, I., Kittler, R., Junqueira, M., Shevchenko, A., Schulz, H., Hubner, N., Doss, M.X., Sachinidis, A., Hescheler, J., Iacone, R., Anastassiadis, K., Stewart, A.F., Pisabarro, M.T., Caldarelli, A., Poser, I., Theis, M. and Buchholz, F. (2009). A genomescale RNAi screen for Oct4 modulators defines a role of the Paf1 complex for embryonic stem cell identity Cell Stem Cell . 4, 403-415.
  6. Ding, L., Paszkowski-Rogacz, M., Nitzsche, A., Slabicki, M.M., Heninger, A.K., de Vries, I., Kittler, R., Junqueira, M., Shevchenko, A., Schulz, H., Hubner, N., Doss, M.X., Sachinidis, A., Hescheler, J., Iacone, R., Anastassiadis, K., Stewart, A.F., Pisabarro, M.T., Caldarelli. A.. Poser, I., Theis, M. and Buchholz, F (2009). A genome-scale RNAi screen for Oct4 modulators defines a role of the Pafl complex for embryonic stem cell identity. Cell Stem Cell. 4, 403-415.
  7. Dothager, R., Flentie, K., Moss, B., Pan, M.H., Kesarwala, A. and Piwnica-Worms, D. (2009). Advances in bioluminescence imaging of live animal models Curr Opin Biotechnol. . 20, 45-53.
  8. Dothager, R., Flentie, K., Moss, B., Pan, M.H., Kesarwala, A., Piwnica-Worms, D. (2009). Advances in bioluminescence imaging of live animal models Curr Opin Biotechnol. 20, 45-53.
  9. Sasportas, L.S., Kasmieh, R., Wakimoto, H., Hingtgen, S., van de Water, J.A., Mohapatra, G., Figueiredo, J.L., Martuza, R.L., Weissleder, R. and Shah, K. (2009). Assessment of therapeutic efficacy and fate of engineered human mesenchymal stem cells for cancer therapy Proc. Natl. Acad. Sci. USA . 106, 4822-4827.
  10. Sasportas, L.S., Kasmieh, R., Wakimoto, H., Hingtgen, S., van de Water, J.A., Mohapatra, G., Figueiredo, J.L., Martuza, R.L., Weissleder, R., Shah, K. (2009). Assessment of therapeutic efficacy and fate of engineered human mesenchymal stem cells for cancer therapy Proc. Natl. Acad. Sci. USA. 106, 4822-4827.
  11. Roda, A., Guardigli, M., Michelini, E., Mirasoli, M. (2009). Bioluminescence in analytical chemistry and in vivo imaging TrAC Trends in Analytical Chemistry. 28, 307-322.
  12. Roda, A., Guardigli, M., Michelini, E. and Mirasoli, M. (2009). Development of a dual luciferase TrAC Trends in Analytical Chemistry . 28, 307-322.
  13. Granneman, J.G., Moore, H.P., Mottillo, E.P. and Zhu, Z.J. (2009). Functional interactions between Mldp (LSDP5) and Abhd5 in the control of intracellular lipid accumulation Biol. Chem. . 284, 3049-3057.
  14. Granneman, J.G., Moore, H.P., Mottillo, E.P. and Zhu, Z. (2009). Functional interactions between Mldp (LSDP5) and Abhd5 in the control of intracellular lipid accumulation. Biol. Chem. 284, 3049-3057.
  15. adr, C.E., Hewett, J.W., Breakefield, X.O. and Tannous, B.A. (2009). The Krüppel-like zinc finger protein Glis3 directly and indirectly activates insulin gene transcription  Nucleic Acids Res. . 37, 2529-2538.
  16. Santos, E., Yes, R., Lee, J., et al (2009). Sensitive in vivo imaging of T cells using membrane-bound gaussia princeps luciferase Nat. Med.. 15, 338-334.
  17. Santos, E., Yeh, R., Lee, J., et al. (2009). Sensitive in vivo imaging of T cells using membrane-bound gaussia princeps luciferase Nat. Med. . 15, 338-344.
  18. Yang, Y., Chang, B.H., Samson, S.L., Li, M.V., Chan, L. (2009). The Kruppel-like zinc finger protein Glis3 directly and indirectly activates insulin gene transcription Nucleic Acids Res.. 37, 2529-2538.
  19. Capul, A.A. and de la Torre, J.C. (2008). A cell-based luciferase assay amenable to high-throughput screening of inhibitors of arenavirus budding Virology . 16-Oct 382, 107- 114.
  20. Capul, A.A. and de la Torre, J.C. (2008). A cell-based luciferase assay amenable to high-throughput screening of inhibitors of arenavirus budding. Virology. 382, 107-114.
  21. Shao, N. and Bock, R. (2008). A codon-optimized luciferase from Gaussia princeps facilitates the in vivo monitoring of gene expression in the model alga Chlamydomonas reinhardtii Curr. Genet. . 53, 381-388.
  22. Shao, N. and Bock, R. (2008). A codon-optimized luciferase from Caussia princeps facilitates the in vivo monitoring of gene expression in the model alga Chlamydomonas reinhardtii. Curr. Genet.. 53, 381-388.
  23. Wurdinger, T., Badr, C., Pike, L., de Kleine, R., Weissleder, R. and Breakefield, X.O. and Tannous, B.A. (2008). A secreted luciferase for ex vivo monitoring of in vivo processes Nat. Methods . 5, 171-173.
  24. Wurdinger, T., Badr, C., Pike, L., de Kleine, R., Weissleder, R., Breakefield, X.O., Tannous, B.A. (2008). A secreted luciferase for ex vivo monitoring of in vivo processes Nat. Methods. 5, 171-173.
  25. Asokan, A., Johnson, J.S., Li, C., Samulski, R.J. (2008). Bioluminescent virion shells; new tools for quantitation of AAV vector dynamics in cells and live animals Gene Ther.. 15, 1618-1622.
  26. Lee, J.Y., Kim, S., Hwang do, W., Jeong, J.M., Chung, J.K., Lee, M.C. and Lee, D.S.J. (2008). Development of a dual-luciferase reporter system for in vivo visualization of MicroRNA biogenesis and posttranscriptional regulation Nucl. Med. . 49, 285-294.
  27. Lee, J.Y., Kim, S., Hwang do, W., Jeong, J.M.,Chung, J.K., Lee, M.C., Lee, D.S.J. (2008). Development of a dual-luciferase reporter system for in vivo visualization of MicroRNA biogenesis and posttranscriptional regulation Nucl. Med.. 49, 285-294.
  28. Ruecker, O., Zillner, K., Groebner-Ferreira, R. and Heitzer, M. (2008). Gaussia-luciferase as a sensitive reporter gene for monitoring promoter activity in the nucleus of the green algaChlamydomonas reinhardtii Mol. Genet. Genomics. . 280153-280162.
  29. Inouye, S. and Sahara, Y. (2008). Identification of two catalytic domains in a luciferase secreted by the copepod Gaussia princeps Biochem Biophys Res Commun. . 4, 96-101.
  30. Michelini, E., Cevenini, L., Mezzanote, L., Ablamsky, D., Southworth, T., Branchini, B. and Roda, A. (2008). Spectral resolved gene technology for multiplexed bioluminescence and high content screening Anal. Chem. . 80, 260-267.
  31. Badr. C.E., Hewett, J.W., Breakefield, X.Q and Tannous, B.A. (2007). A highly sensitive assay for monitoring the secretory pathway and ER stress. PLoS One. 2, e571.
  32. Badr, C.E., Hewett, J.W., Breakefield, X.O. and Tannous, B.A. (2007). A highly sensitive assay for monitoring the secretory pathway and ER stress  PLoS One . 2, e571.
  33. Cheng, G. and Davis, R.E. (2007). An improved and secreted luciferase reporter for schistosomes  Mol. Biochem. Parasitol. . 155, 167-171.
  34. Cheng, G. and Davis, R.E. (2007). An improved and secreted luciferase reporter for schistosomes. Mol. Biochem. Parasitol.. 155, 167-172.
  35. Goerke, A.R. and Loening, A.M., Gambhir, S.S. and Swartz, J.R. (2007). Cell-free metabolic engineering promotes high-level production of bioactive Gaussia princeps luciferase J. Biotechnol. . 128, 705-715.
  36. Morlighem, J.E., Petit, C and Tzertzinis, G. (2007). Determination of silencing potency of synthetic and RNase Ill-generated siRNA using a secreted luciferase assay. Biotechniques. 42, 599-606.
  37. Morlighem, J.E., Petit, C. and Tzertzinis, G. (2007). Determination of silencing potency of synthetic and RNase III-generated siRNA using a secreted luciferase assay Biotechniques . 42, 599-606.
  38. Wu, C., Suzuki-Ogoh, C. and Ohmiya, Y. (2007). Dual-reporter assay using two secreted luciferase genes  Biotechniques . 42, 290-292.
  39. Wu, C., Suzuki-Ogoh, C., Ohmiya, Y. (2007). Dual-reporter assay using two secreted luciferase genes Biotechniques. 42, 290-292.
  40. Venisnik, K.M., Olafsen, T., Gambhir, S.S. and Wu, A.M (2007). Fusion of Gaussia luciferase to an engineered anti-carcinoembryonic antigen (CEA) antibody for in vivo optical imaging  Mol. Imaging Biol. . 9, 267-277.
  41. Venisnik, K.M., Olafsen, T., Gambhir, S.S., Wu, A.M. (2007). Fusion of Gaussia luciferase to an engineered anti-carcinoembryonic antigen (CEA) antibody for in vivo optical imaging Mol. Imaging Biol.. 9, 267-277.
  42. Cheng, G., Cohen, L., Mikhli, C., Jankowska-Anyszka, M., Stepinski, J., Darzynkiewicz, E. and Davis, R.E. (2007). In vivo translation and stability of trans-spliced mRNAs in nematode embryos Mol. Biochem. Parasitol. . 153, 95-106.
  43. Cheng, G., Cohen, L., Mikhli, C, Jankowska-Anyszka, M., Stepinski, J., Darzynkiewicz, E. and Davis, R.E (2007). In vivo translation and stability of trans-spliced mRNAs in nematode embryos. Mol. Biochem. Parasitol.. 153, 95-106.
  44. Hewett, J.W., Tannous, B., Niland. B.P., Nery, F.C., Zeng, J., Li, Y. and Breakefield, X. (2007). Mutant torsinA interferes with protein processing through the secretory pathway in DYT1 dystonia cells. Proc. Natl. Acad. Sci. USA.. 104, 7271-7276.
  45. Hewett, J.W., Tannous, B., Niland, B.P., Nery, F.C., Zeng, J., Li, Y. and Breakefield, X.O. (2007). Mutant torsinA interferes with protein processing through the secretory pathway in DYT1 dystonia cells Proc. Natl. Acad. Sci. USA . 104, 7271-7276.
  46. Knappskog, S., Ravneberg, H., Gjerdrum, C., Trosse, C., Stern, B., and Pryme I.F. (2007). The level of synthesis and secretion of Gaussia princeps luciferase in transfected CHO cells is heavily dependent on the choice of signal peptide J. Biotechnol. 128, 705-715.
  47. Remy, I. and Michnick, S.W. (2006). A highly sensitive protein-protein interaction assay based onGaussia luciferase Nat. Methods . 3, 977-979.
  48. Remy, I. and Michnick, S.W. (2006). A highly sensitive protein-protein interaction assay based onGaussia luciferase Nat. Methods . 3, 977-979.
  49. Remy. I. and Michnick, S.W. (2006). A highly sensitive protein-protein interaction assay based on Gaussia luciferase. Nat. Methods.
  50. Wu, C., Kawasaki, K., Ohgiya.S. and Ohmiya, Y. (2006). Syntheses and evaluation of the bioluminescent activity of (S)-Cypridina luciferin and its analogs Tetrahedron Letters . 47, 753-756.
  51. Wu, C., Kawasaki, K., Ohgiya, S., Ohmiya, Y. (2006). Synthesis and evaluation of the bioluminescent activity of (S)-Cypridina luciferin and its analogs Tetrahedron Letters. 47, 753-756.
  52. Wiles, S., Ferguson, K., Stefanidou, M., Young, D.B. and Robertson, B.D. (2005). Alternative luciferase for monitoring bacterial cells under adverse conditions  Appl. Environ. Microbiol. . 71, PubMedID: 3427-3432
  53. Wiles, S., Ferguson, K., Stefanidou, M., Young, D.B. and Robertson, B.D. (2005). Alternative luciferase for monitoring bacterial cells under adverse conditions. Appl. Environ. Microbiol.. 71, 3427-3432.
  54. Tannous, B.A., Kim, D.E., Fernandez, J.L., Weissleder, R. and Breakefield, X.O. (2005). Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo Mol Ther. . 435-443.
  55. Tannous, B.A., Kim, D.E., Fernandez, J.L., Weissleder, R., Breakefield, X.O. (2005). Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo Mol Ther.. 11, 435-443.
  56. Nakajima, Y., Kobayashi, K., Yamagishi, K., Enomoto, T. and Ohmiya, Y. (2004). cDNA cloning and characterization of a secreted luciferase from the luminous Japanese ostracod, Cypridinanoctiluca Biosci. Biotechnol. Biochem. . 68, 565-570.
  57. Nakajima, Y., Kobayashi, K., Yamagashi, K., Enomoto, T., Ohmiya, Y. (2004). cDNA cloning and characterization of a secreted luciferase from the luminous Japanese ostracod, Cypridina noctiluca Biosci. Biotechnol. Biochem.. 68, 565-570.
  58. Verhaegen, M. and Christopoulos, T.K. (2002). Recombinant Gaussia luciferase. Overexpression, purification, and analytical application of a bioluminescent reporter for DNA hybridization Anal Chem. . 74, 4378-4385.
  59. Verhaegen, M., Christopoulos, T.K. (2002). Recombinant Gaussia luciferase. Overexpression, purification and analytical application of a bioluminescent reporter for DNA hybridization Anal Chem. 74, 4378-4385.

Advantages

  • Naturally Secreted – Used in 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.