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Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals
- Source :
- Nature Methods. 17:852-860
- Publication Year :
- 2020
- Publisher :
- Springer Science and Business Media LLC, 2020.
-
Abstract
- 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. NanoLuc substrates with improved solubility and bioavailability, hydrofurimazine and fluorofurimazine, strongly enhance bioluminescence signals in vivo and enable bright dual-color bioluminescent imaging with AkaLuc and AkaLumine.
- Subjects :
- education.field_of_study
Tumor size
Chemistry
Population
Cell Biology
Biochemistry
Substrate Specificity
Luminescent Proteins
In vivo
Luminescent Measurements
Biophysics
Animals
Substrate specificity
Bioluminescence
Fluorescent protein
Bioluminescence imaging
Luciferase
Furans
Luciferases
education
Molecular Biology
Enzyme Assays
Biotechnology
Subjects
Details
- ISSN :
- 15487105 and 15487091
- Volume :
- 17
- Database :
- OpenAIRE
- Journal :
- Nature Methods
- Accession number :
- edsair.doi.dedup.....1c6045ec5f0ff65f5b482789293d1def