cybLuc: An Effective Aminoluciferin Derivative for Deep Bioluminescence Imaging

Abstract

To enhance the efficiency of firefly luciferase/luciferin bioluminescence imaging, a series of N-cycloalkylaminoluciferins (cyaLucs) were developed by introducing lipophilic N-cycloalkylated substitutions. The experimental results demonstrate that these cyaLucs are effective substrates for native firefly luciferase (Fluc) and can produce elevated bioluminescent signals in vitro, in cellulo, and in vivo. It should be noted that, in animal studies, N-cyclobutylaminoluciferin (cybLuc) at 10 μM (0.1 mL), which is 0.01% of the standard dose of d-luciferin (dLuc) used in mouse imaging, can radiate 20-fold more bioluminescent light than d-luciferin (dLuc) or aminoluciferin (aLuc) at the same concentration. Longer in vivo emission imaging using cybLuc suggests that it can be used for long-time observation. Regarding the mechanism of cybLuc, our cocrystal structure data from firefly luciferase with oxidized cybLuc suggested that oxidized cybLuc fits into the same pocket as oxyluciferin. Most interestingly, our results demonstrate that the sensitivity of cybLuc in brain tumor imaging contributes to its extended application in deep tissues.

Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging

“…This demonstrated that the total light emission and the optimization of imaging conditions and parameters play an important role in the ultimate sensitivity for BLI in vivo. For specific applications in brain that do not require the high resolution provided by NIR-emitting substrates, alternative D-luciferin analogs that produce increased signal have recently been described 36 .…”

Synthesis and bioluminescence of electronically modified and rotationally restricted colour-shifting infraluciferin analogues

“…12,13 Recently, compound 8 and the HCl salt of 5 have shown brighter emission than LH2 at low substrate concentrations. 9,14 Luciferase mutants have also been engineered to give brighter light for some analogues 11,15 including compound 4 under sub-saturating conditions. 16 In 2014, we reported a extended analogue of luciferin infraluciferin (9, iLH2) that exhibited nIR bioluminescence ( max = 706 nm).…”

Anschalten von Festkörperfluoreszenz mit Licht

“…Entlang dieser Argumentationslinie wurden die Aminoderivate des Glühwürmchen‐Luciferins ( 2 in Schema ) kürzlich als potenzielle bioinspirierte Emitter von rotem Licht vorgeschlagen . Obwohl das Potenzial solcher Varianten für die Tiefengewebe‐Bildgebung bereits gezeigt wurde, bleibt die zugrundeliegende Photochemie unklar (detaillierte Informationen zu beschriebenen Aminoluciferinen finden sich in den Hintergrundinformationen, SI). Kürzlich beobachteten wir zufällig, dass Kristalle von N , N ‐Dimethylaminoluciferin 3 (Diagramm 1) ihre Fluoreszenzemission stark erhöhen nachdem sie blauem LED‐Licht ausgesetzt wurden.…”