Evaluation of NanoLuc substrates for bioluminescence imaging of transferred cells in mice.

Details

Serval ID
serval:BIB_66BBF8FB0FE3
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Evaluation of NanoLuc substrates for bioluminescence imaging of transferred cells in mice.
Journal
Journal of photochemistry and photobiology. B, Biology
Author(s)
Gaspar N., Walker J.R., Zambito G., Marella-Panth K., Lowik C., Kirkland T.A., Mezzanotte L.
ISSN
1873-2682 (Electronic)
ISSN-L
1011-1344
Publication state
Published
Issued date
03/2021
Peer-reviewed
Oui
Volume
216
Pages
112128
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
NanoLuc luciferase recently gained popularity due to its small size and superior bioluminescence performance. For in vivo imaging applications, NanoLuc has been limited by its substrate furimazine, which has low solubility and bioavailability. Herein, we compared the performances of recently reported NanoLuc luciferase substrates for in vivo imaging in mice. Two substrates with improved aqueous solubility, hydrofurimazine and fluorofurimazine, were evaluated along with three stabilized O-acetylated furimazine analogues, the hikarazines. All 5 analogues, when tested in vitro, displayed greater signal intensity and reaction duration, in comparison to the standard NanoLuc substrate, furimazine. The two best-performing analogues from the in vitro study were selected for further in vivo testing. The NanoLuc/fluorofurimazine pair demonstrated the highest bioluminescence intensity, post intravenous administration. It was found to be around 9-fold brighter compared to the NanoLuc/furimazine and 11-fold more intense than the NanoLuc/hikarazine-003 pair, with an average of 3-fold higher light emission when the substrate was injected intraperitoneally, in a subcutaneous model. Excitingly, despite the fact that NanoLuc/fluorofurimazine emits mostly blue light, we prove that cells trapped in mice lungs vasculature could be visualised via the NanoLuc/fluorofurimazine pair and compare the results to the AkaLuc/AkaLumine system. Therefore, among the tested analogues, fluorofurimazine enables higher substrate loading and improved optical imaging sensitivity in small animals, upgrading the use of NanoLuc derived bioluminescent systems for deep tissue imaging.
Keywords
AkaLuc, Bioluminescence imaging, Fluorofurimazine, Hikarazines, NanoLuc
Pubmed
Web of science
Open Access
Yes
Create date
08/02/2021 13:18
Last modification date
17/04/2021 5:33
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