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  MOLECULAR IMAGING AND BIOTECHNOLOGY


Minerva Biotecnologica 2009 June;21(2):87-96

language: English

Bioluminescent reporter proteins for multicolor assays

Michelini E. 1, Cevenini L. 1, Mezzanotte L. 1, Roda B. 2, Dolci L. S. 1, Roda A. 1

1 Department of Pharmaceutical Sciences University of Bologna, Bologna, Italy
2 Department of Chemistry “G. Ciamician” University of Bologna, Bologna, Italy


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Bioluminescent (BL) reporter proteins are widely employed in different types of multicolor assays thanks to their peculiar features such as high quantum yield and absence of toxicity when expressed in cells or whole organisms. In particular luciferases are the most common reporter proteins enabling ultrasensitive detection in diverse biotechnological applications. Various bioluminescent proteins have been recently isolated from luminescent organisms and the corresponding genes have been cloned. Wild-type luciferases from fireflies, sea pansies, worms, bacteria and their mutants with improved emission and thermostability are in fact suitable for many applications such as cell-based assays, in vivo imaging, energy transfer-based assays and as new bioluminescent probes in association with quantum dots. Concerning cell-based assays, very few works employing multicolor reporter proteins have been reported in literature. The main bottleneck consists in the spectral unmixing that, when using more than two luciferases emitting at a different wavelength and requiring the same substrate, does not permit to completely separate the signals. Luciferases with red emission (e.g., the red mutants of P. pyralis and L. italica luciferases) may represent the best choice for bioluminescent in vivo imaging since light of wavelengths from 600 to 1100 nm is readily transmitted through tissues.

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