Introducing a new reporter gene, membrane-anchored Cypridina luciferase, for multiplex bioluminescence imaging

Bioluminescence reporter gene imaging is a robust, high-throughput imaging modality that is useful for tracking cells and monitoring biological processes, both in cell culture and in small animals. We introduced and characterized a novel bioluminescence reporter—membrane-anchored Cypridina luciferas...

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Bibliographic Details
Published in:Molecular therapy. Oncolytics 2021-06, Vol.21, p.15-22
Main Authors: Moroz, Maxim A., Zurita, Juan, Moroz, Anna, Nikolov, Ekaterina, Likar, Yury, Dobrenkov, Konstantin, Lee, Jason, Shenker, Larissa, Blasberg, Ronald, Serganova, Inna, Ponomarev, Vladimir
Format: Article
Language:English
Subjects:
bioluminescence imaging
BLI
luciferase reporters
multiplex imaging
Original
reporter gene
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Summary:Bioluminescence reporter gene imaging is a robust, high-throughput imaging modality that is useful for tracking cells and monitoring biological processes, both in cell culture and in small animals. We introduced and characterized a novel bioluminescence reporter—membrane-anchored Cypridina luciferase (maCLuc)—paired with a unique vargulin substrate. This luciferase-substrate pair has no cross-reactivity with established d-luciferin- or coelenterazine-based luciferase reporters. We compare maCLuc with several established luciferase-based reporter systems (firefly, click beetle, Renilla, and Gaussia luciferases), using both in vitro and in vivo models. We demonstrate the different imaging characteristics of these reporter systems, which allow for multiplexed-luciferase imaging of 3 and 4 separate targets concurrently in the same animal within 24 h. The imaging paradigms described here can be directly applied for simultaneous in vivo monitoring of multiple cell populations, the activity of selected signal transduction pathways, or a combination of both constitutive and inducible reporter imaging. [Display omitted] Moroz et al. developed and tested a new reporter system based on a membrane-anchored Cypridina luciferase gene. maCLuc in combination with other established BLI reporter systems provides the opportunity for a new, multiplex imaging paradigm. This paradigm will allow scientists to visualize up to 3 and 4 separate targets concurrently in the same animal.
ISSN:2372-7705
2372-7705
DOI:10.1016/j.omto.2021.03.004