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Publication : Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals.

First Author  Su Y Year  2020
Journal  Nat Methods Volume  17
Issue  8 Pages  852-860
PubMed ID  32661427 Mgi Jnum  J:341167
Mgi Id  MGI:7532761 Doi  10.1038/s41592-020-0889-6
Citation  Su Y, et al. (2020) Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals. Nat Methods 17(8):852-860
abstractText  Sensitive detection of two biological events in vivo has long been a goal in bioluminescence imaging. Antares, a fusion of the luciferase NanoLuc to the orange fluorescent protein CyOFP, has emerged as a bright bioluminescent reporter with orthogonal substrate specificity to firefly luciferase (FLuc) and its derivatives such as AkaLuc. However, the brightness of Antares in mice is limited by the poor solubility and bioavailability of the NanoLuc substrate furimazine. Here, we report a new substrate, hydrofurimazine, whose enhanced aqueous solubility allows delivery of higher doses to mice. In the liver, Antares with hydrofurimazine exhibited similar brightness to AkaLuc with its substrate AkaLumine. Further chemical exploration generated a second substrate, fluorofurimazine, with even higher brightness in vivo. We used Antares with fluorofurimazine to track tumor size and AkaLuc with AkaLumine to visualize CAR-T cells within the same mice, demonstrating the ability to perform two-population imaging with these two luciferase systems.
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