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

Molecular Therapy: Oncolytics

Vladimir Ponomarev¹  Ekaterina Nikolov²  Yury Likar³  Inna Serganova³  Konstantin Dobrenkov⁴  Ronald Blasberg⁴  Maxim A. Moroz⁴  Jason Lee⁴  Juan Zurita⁴  Anna Moroz⁴  Larissa Shenker⁴ 

[1] Molecular Pharmacology and Chemistry Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA;Skolkovo Institute of Science and Technology, Moscow 143026, Russia;Department of Neurology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA;Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA;

关键词: bioluminescence imaging;    BLI;    luciferase reporters;    reporter gene;    multiplex imaging;   

DOI  :  

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown