How to Select Firefly Luciferin Analogues for In Vivo Imaging

Bioluminescence reactions are widely applied in optical in vivo imaging in the life science and medical fields. Such reactions produce light upon the oxidation of a luciferin (substrate) catalyzed by a luciferase (enzyme), and this bioluminescence enables the quantification of tumor cells and gene expression in animal models. Many researchers have developed single-color or multicolor bioluminescence systems based on artificial luciferin analogues and/or luciferase mutants, for application in vivo bioluminescence imaging (BLI). In the current review, we focus on the characteristics of firefly BLI technology and discuss the development of luciferin analogues for high-resolution in vivo BLI. In addition, we discuss the novel luciferin analogues TokeOni and seMpai, which show potential as high-sensitivity in vivo BLI reagents.

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Quantum yield improvement of red-light-emitting firefly luciferin analogues for in vivo bioluminescence imaging

Tetrahedron ◽

10.1016/j.tet.2017.11.051 ◽

2018 ◽

Vol 74 (6) ◽

pp. 652-660 ◽

Cited By ~ 13

Author(s):

Masahiro Kiyama ◽

Satoshi Iwano ◽

Satoshi Otsuka ◽

Shijia W. Lu ◽

Rika Obata ◽

...

Keyword(s):

Quantum Yield ◽

Bioluminescence Imaging ◽

Red Light ◽

Yield Improvement ◽

Light Emitting ◽

Firefly Luciferin ◽

In Vivo Bioluminescence Imaging

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Constructing Firefly Luciferin Bioluminescence Probes for in Vivo Imaging

Organic & Biomolecular Chemistry ◽

10.1039/d1ob01940f ◽

2022 ◽

Author(s):

Xingye Yang ◽

Xiaojun Qin ◽

Huimin Ji ◽

Lupei Du ◽

Minyong Li

Keyword(s):

Real Time ◽

In Vivo Imaging ◽

Bioluminescence Imaging ◽

Biological Systems ◽

Firefly Luciferin ◽

Visual Approach

Bioluminescence imaging (BLI) is a widely applied visual approach for real-time detecting many physiological and pathological processes in a variety of biological systems. Based on the caged strategy, lots of...

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Multicolour In Vivo Bioluminescence Imaging Using a NanoLuc‐Based BRET Reporter in Combination with Firefly Luciferase

Contrast Media & Molecular Imaging ◽

10.1155/2018/2514796 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 6

Author(s):

Arthur Taylor ◽

Jack Sharkey ◽

Antonius Plagge ◽

Bettina Wilm ◽

Patricia Murray

Keyword(s):

Bioluminescence Imaging ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

High Sensitivity ◽

Firefly Luciferase ◽

Cell Populations ◽

Wide Range ◽

In Vivo Bioluminescence Imaging ◽

Multiple Cell

The ability to track the biodistribution and fate of multiple cell populations administered to rodents has the potential to facilitate the understanding of biological processes in a range of fields including regenerative medicine, oncology, and host/pathogen interactions. Bioluminescence imaging is an important tool for achieving this goal, but current protocols rely on systems that have poor sensitivity or require spectral decomposition. Here, we show that a bioluminescence resonance energy transfer reporter (BRET) based on NanoLuc and LSSmOrange in combination with firefly luciferase enables the unambiguous discrimination of two cell populations in vivo with high sensitivity. We insert each of these reporter genes into cells using lentiviral vectors and demonstrate the ability to monitor the cells’ biodistribution under a wide range of administration conditions, including the venous or arterial route, and in different tissues including the brain, liver, kidneys, and tumours. Our protocol allows for the imaging of two cell populations in the same imaging session, facilitating the overlay of the signals and the identification of anatomical positions where they colocalise. Finally, we provide a method for postmortem confirmation of the presence of each cell population in excised organs.

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The Antitumor Effects of Britanin on Hepatocellular Carcinoma Cells and its Real-Time Evaluation by In Vivo Bioluminescence Imaging

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200227092623 ◽

2020 ◽

Vol 20 (9) ◽

pp. 1147-1156

Author(s):

Hanrui Li ◽

GeTao Du ◽

Lu Yang ◽

Liaojun Pang ◽

Yonghua Zhan

Keyword(s):

Hepatocellular Carcinoma ◽

Western Blot ◽

Blot Analysis ◽

Tumor Size ◽

Hepg2 Cells ◽

Western Blot Analysis ◽

Bioluminescence Imaging ◽

Antitumor Effects ◽

In Vivo Bioluminescence Imaging

Background: Hepatocellular carcinoma is cancer with many new cases and the highest mortality rate. Chemotherapy is the most commonly used method for the clinical treatment of hepatocellular carcinoma. Natural products have become clinically important chemotherapeutic drugs due to their great potential for pharmacological development. Many sesquiterpene lactone compounds have been proven to have antitumor effects on hepatocellular carcinoma. Objective: Britanin is a sesquiterpene lactone compound that can be considered for the treatment of hepatocellular carcinoma. The present study aimed to investigate the antitumor effect of britanin. Methods: BEL 7402 and HepG2 cells were used to study the cytotoxicity and antitumor effects of britanin. Preliminary studies on the nuclear factor kappa B pathway were conducted by western blot analysis. A BEL 7402-luc subcutaneous tumor model was established for the in vivo antitumor studies of britanin. In vivo bioluminescence imaging was conducted to monitor changes in tumor size. Results: The results of the cytotoxicity analysis showed that the IC50 values for britanin in BEL 7402 and HepG2 cells were 2.702μM and 6.006μM, respectively. The results of the colony formation demonstrated that the number of cells in a colony was reduced significantly after britanin treatment. And the results of transwell migration assays showed that the migration ability of tumor cells was significantly weakened after treatment with britanin. Tumor size measurements and staining results showed that tumor size was inhibited after britanin treatment. The western blot analysis results showed the inhibition of p65 protein expression and reduced the ratio of Bcl-2/Bax after treatment. Conclusion: A series of in vitro and in vivo experiments demonstrated that britanin had good antitumor effects and provided an option for hepatocellular carcinoma treatment.

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