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S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells

A unique combinatorial bioluminescence (BL) imaging system was developed for determining molecular events in mammalian cells with various colors and BL intensity patterns. This imaging system consists of one or multiple reporter luciferases and a series of novel coelenterazine (CTZ) analogues named...

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Autores principales: Kamiya, Genta, Kitada, Nobuo, Furuta, Tadaomi, Hirano, Takashi, Maki, Shojiro A., Kim, Sung-Bae
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9865520/
https://www.ncbi.nlm.nih.gov/pubmed/36674934
http://dx.doi.org/10.3390/ijms24021420
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author Kamiya, Genta
Kitada, Nobuo
Furuta, Tadaomi
Hirano, Takashi
Maki, Shojiro A.
Kim, Sung-Bae
author_facet Kamiya, Genta
Kitada, Nobuo
Furuta, Tadaomi
Hirano, Takashi
Maki, Shojiro A.
Kim, Sung-Bae
author_sort Kamiya, Genta
collection PubMed
description A unique combinatorial bioluminescence (BL) imaging system was developed for determining molecular events in mammalian cells with various colors and BL intensity patterns. This imaging system consists of one or multiple reporter luciferases and a series of novel coelenterazine (CTZ) analogues named “S-series”. For this study, ten kinds of novel S-series CTZ analogues were synthesized and characterized concerning the BL intensities, spectra, colors, and specificity of various marine luciferases. The characterization revealed that the S-series CTZ analogues luminesce with blue-to-orange-colored BL spectra with marine luciferases, where the most red-shifted BL spectrum peaked at 583 nm. The colors completed a visible light color palette with those of our precedent C-series CTZ analogues. The synthesized substrates S1, S5, S6, and S7 were found to have a unique specificity with marine luciferases, such as R86SG, NanoLuc (shortly, NLuc), and ALuc16. They collectively showed unique BL intensity patterns to identify the marine luciferases together with colors. The marine luciferases, R86SG, NLuc, and ALuc16, were multiplexed into multi-reporter systems, the signals of which were quantitatively unmixed with the specific substrates. When the utility was applied to a single-chain molecular strain probe, the imaging system simultaneously reported three different optical indexes for a ligand, i.e., unique BL intensity and color patterns for identifying the reporters, together with the ligand-specific fold intensities in mammalian cells. This study directs a new combinatorial BL imaging system to specific image molecular events in mammalian cells with multiple optical indexes.
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spelling pubmed-98655202023-01-22 S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells Kamiya, Genta Kitada, Nobuo Furuta, Tadaomi Hirano, Takashi Maki, Shojiro A. Kim, Sung-Bae Int J Mol Sci Article A unique combinatorial bioluminescence (BL) imaging system was developed for determining molecular events in mammalian cells with various colors and BL intensity patterns. This imaging system consists of one or multiple reporter luciferases and a series of novel coelenterazine (CTZ) analogues named “S-series”. For this study, ten kinds of novel S-series CTZ analogues were synthesized and characterized concerning the BL intensities, spectra, colors, and specificity of various marine luciferases. The characterization revealed that the S-series CTZ analogues luminesce with blue-to-orange-colored BL spectra with marine luciferases, where the most red-shifted BL spectrum peaked at 583 nm. The colors completed a visible light color palette with those of our precedent C-series CTZ analogues. The synthesized substrates S1, S5, S6, and S7 were found to have a unique specificity with marine luciferases, such as R86SG, NanoLuc (shortly, NLuc), and ALuc16. They collectively showed unique BL intensity patterns to identify the marine luciferases together with colors. The marine luciferases, R86SG, NLuc, and ALuc16, were multiplexed into multi-reporter systems, the signals of which were quantitatively unmixed with the specific substrates. When the utility was applied to a single-chain molecular strain probe, the imaging system simultaneously reported three different optical indexes for a ligand, i.e., unique BL intensity and color patterns for identifying the reporters, together with the ligand-specific fold intensities in mammalian cells. This study directs a new combinatorial BL imaging system to specific image molecular events in mammalian cells with multiple optical indexes. MDPI 2023-01-11 /pmc/articles/PMC9865520/ /pubmed/36674934 http://dx.doi.org/10.3390/ijms24021420 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kamiya, Genta
Kitada, Nobuo
Furuta, Tadaomi
Hirano, Takashi
Maki, Shojiro A.
Kim, Sung-Bae
S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title_full S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title_fullStr S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title_full_unstemmed S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title_short S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells
title_sort s-series coelenterazine-driven combinatorial bioluminescence imaging systems for mammalian cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9865520/
https://www.ncbi.nlm.nih.gov/pubmed/36674934
http://dx.doi.org/10.3390/ijms24021420
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