A Series of Furimazine Derivatives for Sustained Live-Cell Bioluminescence Imaging and Application to the Monitoring of Myogenesis at the Single-Cell Level

Orioka, Mariko; Eguchi, Masatoshi; Mizui, Yuki; Ikeda, Yoshikazu; Sakama, Akihiro; Li, Qiaojing; Yoshimura, Hideyuki; Ozawa, Toshio; Citterio, Daniel; Hiruta, Yuki

doi:10.1021/acs.bioconjchem.2c00035

Cited by 11 publications

(14 citation statements)

References 61 publications

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“…One drawback of using NLuc may be substrate instability compared to that of other luciferases. However, stable substrates for NLuc have been recently developed, , allowing long-term luminescence imaging. Consequently, the usage of bioluminescence will expand the possibility of multimodal bioimaging in which different molecules, including RNAs and related proteins, are observed in the same cells.…”

Section: Resultsmentioning

confidence: 99%

Split Luciferase-Fragment Reconstitution for Unveiling RNA Localization and Dynamics in Live Cells

Eguchi,

Yoshimura,

Ueda

et al. 2023

ACS Sens.

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The intracellular distribution and dynamics of RNAs play pivotal roles in various physiological phenomena. The ability to monitor the amount and localization of endogenous RNAs in living cells allows for elucidating the mechanisms of various intracellular events. Protein-based fluorescent RNA probes are now widely used to visualize and analyze RNAs in living cells. However, continuously monitoring the temporal changes in RNA localization and dynamics in living cells is challenging. In this study, we developed a bioluminescent probe for spatiotemporal monitoring of RNAs in living cells by using a split-luciferase reconstitution technique. The probe consists of split fragments of a bioluminescent protein, NanoLuc, connected with RNA-binding protein domains generated from a custom-made mutation of a PUM-HD. The probe showed rapid luminescence intensity changes in response to an increase or decrease in the amount of a target RNA in vitro. In live-cell imaging, temporal alteration of the intracellular distribution of endogenous β-actin mRNA was visualized in response to extracellular stimulation. Furthermore, the application of the probe to the visualization of the specific localization of β-actin mRNA in primary hippocampal neurons was conducted. These results demonstrate the capability of the bioluminescent RNA probe to monitor the changes in localization, dynamics, and the amount of target RNA in living cells.

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Section: Resultsmentioning

confidence: 99%

Split Luciferase-Fragment Reconstitution for Unveiling RNA Localization and Dynamics in Live Cells

Eguchi,

Yoshimura,

Ueda

et al. 2023

ACS Sens.

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“…Of note, several other studies developed C-3 caged CTZ or FRZ analogs, some of which also slowed bioluminescence decays in living cells 50 , 51 . Particularly, a substrate manufactured by Promega Corporation, namely ViviRen, was shown to improve the light output of Renilla luciferase in live mice bearing brain tumors compared to the equimolar amount of CTZ 30 .…”

Section: Discussionmentioning

confidence: 99%

A luciferase prosubstrate and a red bioluminescent calcium indicator for imaging neuronal activity in mice

Tian

Zhang

et al. 2022

Nat Commun

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Although fluorescent indicators have been broadly utilized for monitoring bioactivities, fluorescence imaging, when applied to mammals, is limited to superficial targets or requires invasive surgical procedures. Thus, there is emerging interest in developing bioluminescent indicators for noninvasive mammalian imaging. Bioluminescence imaging (BLI) of neuronal activity is highly desired but hindered by insufficient photons needed to digitalize fast brain activities. In this work, we develop a luciferase prosubstrate deliverable at an increased dose and activated in vivo by nonspecific esterase. We further engineer a bright, bioluminescent indicator with robust responsiveness to calcium ions (Ca2+) and appreciable emission above 600 nm. Integration of these advantageous components enables the imaging of the activity of neuronal ensembles in awake mice minimally invasively with excellent signal-to-background and subsecond temporal resolution. This study thus establishes a paradigm for studying brain function in health and disease.

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“…[25][26][27] Novel coelenterazine-type bioluminescent probes pioneered by the Li group were also found sensitive and selective in detecting NTR (Figure 1b). [28] Although controlling the release of the bulky ester from furimazine allows bioluminescence imaging of the hydrolase in living mammalian cells, [29][30] no study has adopted the NLuc-furimazine luminescent system into the detection of nitroreductase in bacteria. Shed on the light of their notable contrast in bioluminescence, [31][32] we introduced a caged group on the C-3 position to report the development of the first furimazine probe for bioluminescent detection of NTR in bacteria.…”

Section: Background and Originality Contentmentioning

confidence: 99%

An Efficient Probe for Bacterial Nitroreductase Imaging and Detection Based on NanoLuc‐Furimazine Bioluminescent Pair

Shi,

Wang,

et al. 2024

Chin. J. Chem.

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Comprehensive SummaryThe detection of critical endogenous species, such as bacteria in microenvironments in the body, requires better imaging tools for visualization and monitoring of biological events. Bioluminescence imaging is the most popular strategy for obtaining real‐time in living cells and organisms. Herein, we introduced a nitroaryl group on the C‐3 position and a hydroxy group at the C‐6 phenyl ring on furimazine to report the first bioluminescent probe (7) based on NanoLuc‐furimazine bioluminescent pair for the detection of nitroreductase in bacteria. The probe, which possessed up to 560‐fold intensity increase with a low detection limit of 16 ng/mL of nitroreductase, has the most efficient uncage efficiency in comparison with other bioluminescent congeners, thus enabling highly selective and sensitive visualization of NTR activity in a panel of clinical priority pathogens. Additionally, imaging of the recombinant strain as well as the NTR from mouse feces indicated the potential of this probe in the application of different mouse disease models.

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A Series of Furimazine Derivatives for Sustained Live-Cell Bioluminescence Imaging and Application to the Monitoring of Myogenesis at the Single-Cell Level

Cited by 11 publications

References 61 publications

Split Luciferase-Fragment Reconstitution for Unveiling RNA Localization and Dynamics in Live Cells

Split Luciferase-Fragment Reconstitution for Unveiling RNA Localization and Dynamics in Live Cells

A luciferase prosubstrate and a red bioluminescent calcium indicator for imaging neuronal activity in mice

An Efficient Probe for Bacterial Nitroreductase Imaging and Detection Based on NanoLuc‐Furimazine Bioluminescent Pair

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