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Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals.
Nature Methods ( IF 48.0 ) Pub Date : 2020-07-13 , DOI: 10.1038/s41592-020-0889-6
Yichi Su 1, 2 , Joel R Walker 3 , Yunhee Park 1, 2 , Thomas P Smith 3 , Lan Xiang Liu 1, 2 , Mary P Hall 4 , Louai Labanieh 2 , Robin Hurst 4 , David C Wang 1, 5 , Lance P Encell 4 , Namdoo Kim 1, 2, 6 , Feijie Zhang 7, 8 , Mark A Kay 7, 8 , Kerriann M Casey 9 , Robbie G Majzner 7, 10 , Jennifer R Cochran 2 , Crystal L Mackall 7, 11 , Thomas A Kirkland 3 , Michael Z Lin 1, 2, 7
Affiliation  

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.



中文翻译:

新型NanoLuc底物可在动物体内进行明亮的两种群生物发光成像。

体内两种生物事件的灵敏检测一直是生物发光成像的目标。Antares是萤光素酶NanoLuc与橙色荧光蛋白CyOFP的融合体,已经成为一种明亮的生物发光报告基因,对萤火虫萤光素酶(FLuc)及其衍生物(如AkaLuc)具有正交的底物特异性。但是,小鼠中Antares的亮度受到NanoLuc底物呋喃嗪的不良溶解度和生物利用度的限制。在这里,我们报告了一种新的底物氢呋喃嗪,其水溶性增强使得可以向小鼠递送更高剂量。在肝脏中,带有氢呋喃嗪的安塔雷(Antares)的亮度与带有底物AkaLumine的AkaLuc相似。进一步的化学探索产生了第二种底物,氟呋喃嗪,在体内具有更高的亮度。

更新日期:2020-07-13
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