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Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Biosensors".

Deadline for manuscript submissions: closed (15 January 2024) | Viewed by 8209

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Dr. Sung-Bae Kim

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Guest Editor

Environmental Management Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba West, 16-1, Onogawa, Tsukuba, Ibaraki 305-8569, Japan
Interests: bioluminescence; luciferase; single-chain bioluminescence probe; molecular imaging; bioassay; BRET

Prof. Dr. Ramasamy Paulmurugan

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Guest Editor

Department of Radiology, Molecular Imaging Program at Stanford, Canary Centre for Cancer Early Detection, Bio-X Program, Stanford University School of Medicine, Palo Alto, CA 94304, USA
Interests: molecular imaging; cancer therapy; vaccine development
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Special Issue Information

Dear Colleagues,

Bioluminescence (BL) is a cold body radiation of light generated by a luciferase-catalyzed oxidation of a luciferin in living organisms. To date, BL has been rapidly emerging in the applications to bioassays and molecular imaging. It has been greatly harnessed by its distinctive advantages, such as the long dynamic range, high signal-to-noise ratios, and no need for external light source for excitation. Reviewing the recent BL studies, a great diversity has been witnessed in the study landscape from the basic to applied sciences: e.g., organic synthesis of luciferins, genetic mutations of luciferases, investigations on the light-emitting mechanisms, development of new bioluminescent probes, and even imaging of molecular events in animal models. This Special Issue of Sensors is intended to provide a communication platform for the latest developments in basic and applied science applications in BL studies, and showcases the bioanalytical utilities in cells and animal models. This Special Issue opens the opportunity for communication and welcomes the broad range of research articles.

Dr. Sung-bae Kim
Prof. Dr. Ramasamy Paulmurugan
Guest Editors

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Published Papers (6 papers)

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Research

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12 pages, 3836 KiB

Open AccessArticle

Luminescent Bacteria as Bioindicators in Screening and Selection of Enzymes Detoxifying Various Mycotoxins

by Elena Efremenko, Ilya Lyagin, Nikolay Stepanov, Olga Senko, Olga Maslova, Aysel Aslanli and Natalia Ugarova

Sensors 2024, 24(3), 763; https://doi.org/10.3390/s24030763 - 24 Jan 2024

Viewed by 655

Abstract

Interest in enzymes capable of neutralizing various mycotoxins is quite high. The methods used for the screening and selection of enzymes that catalyze the detoxification of mycotoxins should be sensitive and fast. However toxic compounds can be generated under the action of such enzymes. Thus, the assessment of the overall reduction in the toxic properties of reaction media towards bioluminescent bacteria seems to be the most reasonable control method allowing a quick search for the effective enzymatic biocatalysts. The influence of a wide range of mycotoxins and glucanases, which hydrolyze toxins with different chemical structures, on the analytical characteristics of luminescent photobacteria as a biosensing element has been studied. Different glucanases (β-glucosidase and endoglucanase) were initially selected for reactions with 10 mycotoxins based on the results of molecular docking which was performed in silico with 20 mycotoxins. Finally, the biorecognizing luminescent cells were used to estimate the residual toxicity of reaction media with mycotoxins after their interaction with enzymes. The notable non-catalytic decrease in toxicity of media containing deoxynivalenol was revealed with luminous cells for both types of tested glucanases, whereas β-glucosidase provided a significant catalytic detoxification of media with aflatoxin B₂ and zearalenone at pH 6.0. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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0 pages, 28203 KiB

Open AccessArticle

Triple Reporter Assay: A Non-Overlapping Luciferase Assay for the Measurement of Complex Macromolecular Regulation in Cancer Cells Using a New Mushroom Luciferase–Luciferin Pair

by Aaiyas Mujawar, Pratham Phadte, Ksenia A. Palkina, Nadezhda M. Markina, Ameena Mohammad, Bhushan L. Thakur, Karen S. Sarkisyan, Anastasia V. Balakireva, Pritha Ray, Ilia Yampolsky and Abhijit De

Sensors 2023, 23(17), 7313; https://doi.org/10.3390/s23177313 - 22 Aug 2023

Viewed by 1649

Abstract

This study demonstrates the development of a humanized luciferase imaging reporter based on a recently discovered mushroom luciferase (Luz) from Neonothopanus nambi. In vitro and in vivo assessments showed that human-codon-optimized Luz (hLuz) has significantly higher activity than native Luz in various cancer cell types. The potential of hLuz in non-invasive bioluminescence imaging was demonstrated by human tumor xenografts subcutaneously and by the orthotopic lungs xenograft in immunocompromised mice. Luz enzyme or its unique 3OH-hispidin substrate was found to be non-cross-reacting with commonly used luciferase reporters such as Firefly (FLuc2), Renilla (RLuc), or nano-luciferase (NLuc). Based on this feature, a non-overlapping, multiplex luciferase assay using hLuz was envisioned to surpass the limitation of dual reporter assay. Multiplex reporter functionality was demonstrated by designing a new sensor construct to measure the NF-κB transcriptional activity using hLuz and utilized in conjunction with two available constructs, p53-NLuc and PIK3CA promoter-FLuc2. By expressing these constructs in the A2780 cell line, we unveiled a complex macromolecular regulation of high relevance in ovarian cancer. The assays performed elucidated the direct regulatory action of p53 or NF-κB on the PIK3CA promoter. However, only the multiplexed assessment revealed further complexities as stabilized p53 expression attenuates NF-κB transcriptional activity and thereby indirectly influences its regulation on the PIK3CA gene. Thus, this study suggests the importance of live cell multiplexed measurement of gene regulatory function using more than two luciferases to address more realistic situations in disease biology. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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16 pages, 3933 KiB

Open AccessArticle

Creation of Artificial Luciferase 60s from Sequential Insights and Their Applications to Bioassays

by Sung-Bae Kim, Tadaomi Furuta, Nobuo Kitada and Shojiro A. Maki

Sensors 2023, 23(14), 6376; https://doi.org/10.3390/s23146376 - 13 Jul 2023

Viewed by 907

Abstract

In this study, a series of new artificial luciferases (ALucs) was created using sequential insights on missing peptide blocks, which were revealed using the alignment of existing ALuc sequences. Through compensating for the missing peptide blocks in the alignment, 10 sibling sequences were artificially fabricated and named from ALuc55 to ALuc68. The phylogenetic analysis showed that the new ALucs formed an independent branch that was genetically isolated from other natural marine luciferases. The new ALucs successfully survived and luminesced with native coelenterazine (nCTZ) and its analogs in living mammalian cells. The results showed that the bioluminescence (BL) intensities of the ALucs were interestingly proportional to the length of the appended peptide blocks. The computational modeling revealed that the appended peptide blocks created a flexible region near the active site, potentially modulating the enzymatic activities. The new ALucs generated various colors with maximally approximately 90 nm redshifted BL spectra in orange upon reaction with the authors’ previously reported 1- and 2-series coelenterazine analogs. The utilities of the new ALucs in bioassays were demonstrated through the construction of single-chain molecular strain probes and protein fragment complementation assay (PCA) probes. The success of this study can guide new insights into how we can engineer and functionalize marine luciferases to expand the toolbox of optical readouts for bioassays and molecular imaging. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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15 pages, 1753 KiB

Open AccessArticle

Coelenterazine Indicators for the Specific Imaging of Human and Bovine Serum Albumins

by Sung-Bae Kim, Genta Kamiya, Tadaomi Furuta, Nobuo Kitada and Shojiro A. Maki

Sensors 2023, 23(13), 6020; https://doi.org/10.3390/s23136020 - 29 Jun 2023

Cited by 1 | Viewed by 1064

Abstract

Albumin assays in serum are important for the prognostic assessment of many life-threatening diseases, such as heart failure, liver disease, malnutrition, inflammatory bowel disease, infections, and kidney disease. In this study, synthetic coelenterazine (CTZ) indicators are developed to quantitatively illuminate human and bovine serum albumins (HSA and BSA) with high specificity. Their functional groups were chemically modified to specifically emit luminescence with HSA and BSA. The CTZ indicators were characterized by assaying the most abundant serum proteins and found that the CTZ indicators S6 and S6h were highly specific to HSA and BSA, respectively. Their colors were dramatically converted from blue, peaked at 480 nm, to yellowish green, peaked at 535 nm, according to the HSA–BSA mixing ratios, wherein the origins and mixing levels of the albumins can be easily determined by their colors and peak positions. The kinetic properties of HSA and BSA were investigated in detail, confirming that the serum albumins catalyze the CTZ indicators, which act as pseudo-luciferases. The catalytic reactions were efficiently inhibited by specific inhibitors, blocking the drug-binding sites I and II of HSA and BSA. Finally, the utility of the CTZ indicators was demonstrated through a quantitative imaging of the real fetal bovine serum (FBS). This study is the first example to show that the CTZ indicators specify HSA and BSA with different colors. This study contributes to the expansion of the toolbox of optical indicators, which efficiently assays serum proteins in physiological samples. Considering that these CTZ indicators immediately report quantitative optical signals with high specificity, they provide solutions to conventional technical hurdles on point-of-care assays of serum albumins. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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16 pages, 4611 KiB

Open AccessArticle

Bright Molecular Strain Probe Templates for Reporting Protein–Protein Interactions

by Sung-Bae Kim, Tadaomi Furuta, Genta Kamiya, Nobuo Kitada, Ramasamy Paulmurugan and Shojiro A. Maki

Sensors 2023, 23(7), 3498; https://doi.org/10.3390/s23073498 - 27 Mar 2023

Viewed by 1496

Abstract

Imaging protein–protein interactions (PPIs) is a hot topic in molecular medicine in the postgenomic sequencing era. In the present study, we report bright and highly sensitive single-chain molecular strain probe templates which embed full-length Renilla luciferase 8.6-535SG (RLuc86SG) or Artificial luciferase 49 (ALuc49) as reporters. These reporters were deployed between FKBP-rapamycin binding domain (FRB) and FK506-binding protein (FKBP) as a PPI model. This unique molecular design was conceptualized to exploit molecular strains of the sandwiched reporters appended by rapamycin-triggered intramolecular PPIs. The ligand-sensing properties of the templates were maximized by interface truncations and substrate modulation. The highest fold intensities, 9.4 and 16.6, of the templates were accomplished with RLuc86SG and ALuc49, respectively. The spectra of the templates, according to substrates, revealed that the colors are tunable to blue, green, and yellow. The putative substrate-binding chemistry and the working mechanisms of the probes were computationally modeled in the presence or absence of rapamycin. Considering that the molecular strain probe templates are applicable to other PPI models, the present approach would broaden the scope of the bioassay toolbox, which harnesses the privilege of luciferase reporters and the unique concept of the molecular strain probes into bioassays and molecular imaging. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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15 pages, 1967 KiB

Open AccessReview

Illuminating Progress: The Contribution of Bioluminescence to Sustainable Development Goal 6—Clean Water and Sanitation—Of the United Nations 2030 Agenda

by Denise Gregucci, Faisal Nazir, Maria Maddalena Calabretta and Elisa Michelini

Sensors 2023, 23(16), 7244; https://doi.org/10.3390/s23167244 - 18 Aug 2023

Viewed by 1649

Abstract

The United Nations Agenda 2030 Sustainable Development Goal 6 (SDG 6) aims at ensuring the availability and sustainable management of water and sanitation. The routine monitoring of water contaminants requires accurate and rapid analytical techniques. Laboratory analyses and conventional methods of field sampling still require considerable labor and time with highly trained personnel and transport to a central facility with sophisticated equipment, which renders routine monitoring cumbersome, time-consuming, and costly. Moreover, these methods do not provide information about the actual toxicity of water, which is crucial for characterizing complex samples, such as urban wastewater and stormwater runoff. The unique properties of bioluminescence (BL) offer innovative approaches for developing advanced tools and technologies for holistic water monitoring. BL biosensors offer a promising solution by combining the natural BL phenomenon with cutting-edge technologies. This review provides an overview of the recent advances and significant contributions of BL to SDG 6, focusing attention on the potential use of the BL-based sensing platforms for advancing water management practices, protecting ecosystems, and ensuring the well-being of communities. Full article

(This article belongs to the Special Issue Bioluminescence Imaging and Sensing: Applications in Basic and Applied Sciences)

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