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Advances in Luciferase
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Advances in Luciferase

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: closed (30 August 2024) | Viewed by 12760

Special Issue Editor

Dr. Yoshhiro Ohmiya

E-Mail Website
Guest Editor
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan
Interests: bioluminescence; luciferase

Special Issue Information

Dear Colleagues,

Bioluminescence, the production of light without heat by living organisms, has elicited the interest of scientists for a long time. The biological functions of bioluminescence are thought to involve defense against predators, assistance in predation, communication in reproduction, and metabolic as well as biochemical pathway byproduct formation in organisms. On the other hand, bioluminescence is the emission of light based on the luciferin–luciferase enzymatic reaction in living organisms. Light signals from bioluminescence reactions are available for bioanalysis and bioreporters for gene expression, in vitro, in vivo, and ex vivo bioimaging, immunoassays, and other applications. Additionally, based on the biosynthesis of luciferin, beetle bioluminescence can detect pesticides, or fungus luminescence can produce auto-luminescence in plants; therefore, the possibilities of bioluminescence system could be infinite. This open access Special Issue will bring together original research and review articles on basic applications of bioluminescence systems and on new knowledge of luciferase as well as luciferin. It highlights new discoveries, approaches, and technical developments in bioluminescence research. The main feature of this Special Issue is to provide open source sharing of significant work.

Topics of this Special Issue include, but are not limited to, the following:

  • Applications of bioluminescence systems, including gene expression analysis, in vitro, in vivo, and ex vivo bioimaging, immunoassays, etc.;
  • Cloning and characterization of new luciferase;
  • Determining the biosynthesis pathway of luciferin;
  • Identifying new luciferin from bioluminescence organisms;
  • Modification and characterization of luciferase or luciferin;
  • Molecular mechanisms of luciferin–luciferase reactions.

Dr. Yoshhiro Ohmiya
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • bioluminescence
  • bioimaging
  • biosynthesis
  • color modulation evolution
  • immunoassay
  • immunohistrochemistry
  • light signal
  • luciferase
  • luciferin
  • luciferase modification
  • luciferin biosynthesis
  • luciferin modification
  • molecular mechnism
  • screening
  • structure
  • toxycological test

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Published Papers (6 papers)

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Research

Jump to: Review, Other

14 pages, 12752 KiB  
Article
Establishment of Translational Luciferase-Based Cancer Models to Evaluate Antitumoral Therapies
by Martin R. Ramos-Gonzalez, Nagabhishek Sirpu Natesh, Satyanarayana Rachagani, James Amos-Landgraf, Haval Shirwan, Esma S. Yolcu and Jorge G. Gomez-Gutierrez
Int. J. Mol. Sci. 2024, 25(19), 10418; https://doi.org/10.3390/ijms251910418 - 27 Sep 2024
Cited by 2 | Viewed by 1994
Abstract
Luciferase (luc) bioluminescence (BL) is the most used light-emitting protein that has been engineered to be expressed in multiple cancer cell lines, allowing for the detection of tumor nodules in vivo as it can penetrate most tissues. The goal of this study was [...] Read more.
Luciferase (luc) bioluminescence (BL) is the most used light-emitting protein that has been engineered to be expressed in multiple cancer cell lines, allowing for the detection of tumor nodules in vivo as it can penetrate most tissues. The goal of this study was to develop an oncolytic adenovirus (OAd)-resistant human triple-negative breast cancer (TNBC) that could express luciferase. Thus, when combining an OAd with chemotherapies or targeted therapies, we would be able to monitor the ability of these compounds to enhance OAd antitumor efficacy using BL in real time. The TNBC cell line HCC1937 was stably transfected with the plasmid pGL4.50[luc2/CMV/Hygro] (HCC1937/luc2). Once established, HCC1937/luc2 was orthotopically implanted in the 4th mammary gland fat pad of NSG (non-obese diabetic severe combined immunodeficiency disease gamma) female mice. Bioluminescence imaging (BLI) revealed that the HCC1937/luc2 cell line developed orthotopic breast tumor and lung metastasis over time. However, the integration of luc plasmid modified the HCC1937 phenotype, making HCC1937/luc2 more sensitive to OAdmCherry compared to the parental cell line and blunting the interferon (IFN) antiviral response. Testing two additional luc cell lines revealed that this was not a universal response; however, proper controls would need to be evaluated, as the integration of luciferase could affect the cells’ response to different treatments. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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17 pages, 3690 KiB  
Article
A Novel Luciferase-Based Reporter Gene Technology for Simultaneous Optical and Radionuclide Imaging of Cells
by Natasa Gaspar, Maryana Handula, Marcus C. M. Stroet, Kranthi Marella-Panth, Joost Haeck, Thomas A. Kirkland, Mary P. Hall, Lance P. Encell, Simone Dalm, Clemens Lowik, Yann Seimbille and Laura Mezzanotte
Int. J. Mol. Sci. 2024, 25(15), 8206; https://doi.org/10.3390/ijms25158206 - 27 Jul 2024
Viewed by 3463
Abstract
Multimodality reporter gene imaging combines the sensitivity, resolution and translational potential of two or more signals. The approach has not been widely adopted by the animal imaging community, mainly because its utility in this area is unproven. We developed a new complementation-based reporter [...] Read more.
Multimodality reporter gene imaging combines the sensitivity, resolution and translational potential of two or more signals. The approach has not been widely adopted by the animal imaging community, mainly because its utility in this area is unproven. We developed a new complementation-based reporter gene system where the large component of split NanoLuc luciferase (LgBiT) presented on the surface of cells (TM-LgBiT) interacts with a radiotracer consisting of the high-affinity complementary HiBiT peptide labeled with a radionuclide. Radiotracer uptake could be imaged in mice using SPECT/CT and bioluminescence within two hours of implanting reporter-gene-expressing cells. Imaging data were validated by ex vivo biodistribution studies. Following the demonstration of complementation between the TM-LgBiT protein and HiBiT radiotracer, we validated the use of the technology in the highly specific in vivo multimodal imaging of cells. These findings highlight the potential of this new approach to facilitate the advancement of cell and gene therapies from bench to clinic. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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11 pages, 1640 KiB  
Article
Serum Amyloid A3 Promoter-Luciferase Reporter Mice Are Useful for Early Drug-Induced Nephrotoxicity Detection
by Ayane Kudo, Haruka Osedo, Rahmawati Aisyah, Nao Yazawa, Tolulope Peter Saliu, Kenshu Miyata, Thanutchaporn Kumrungsee and Noriyuki Yanaka
Int. J. Mol. Sci. 2024, 25(10), 5124; https://doi.org/10.3390/ijms25105124 - 8 May 2024
Cited by 1 | Viewed by 1243
Abstract
Early detection of drug-induced kidney injury is essential for drug development. In this study, multiple low-dose aristolochic acid (AA) and cisplatin (Cis) injections increased renal mRNA levels of inflammation, fibrosis, and renal tubule injury markers. We applied a serum amyloid A3 (Saa3) promoter-driven [...] Read more.
Early detection of drug-induced kidney injury is essential for drug development. In this study, multiple low-dose aristolochic acid (AA) and cisplatin (Cis) injections increased renal mRNA levels of inflammation, fibrosis, and renal tubule injury markers. We applied a serum amyloid A3 (Saa3) promoter-driven luciferase reporter (Saa3 promoter-luc mice) to these two tubulointerstitial nephritis models and performed in vivo bioluminescence imaging to monitor early renal pathologies. The bioluminescent signals from renal tissues with AA or CIS injections were stronger than those from normal kidney tissues obtained from normal mice. To verify whether the visualized bioluminescence signal was specifically generated by the injured kidney, we performed in vivo bioluminescence analysis after opening the stomachs of Saa3 promoter-luc mice, and the Saa3-mediated bioluminescent signal was specifically detected in the injured kidney. This study showed that Saa3 promoter activity is a potent non-invasive indicator for the early detection of drug-induced nephrotoxicity. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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15 pages, 3451 KiB  
Article
Multiplex Detection of Fluorescent Chemokine Binding to CXC Chemokine Receptors by NanoBRET
by Justyna M. Adamska, Spyridon Leftheriotis, Reggie Bosma, Henry F. Vischer and Rob Leurs
Int. J. Mol. Sci. 2024, 25(9), 5018; https://doi.org/10.3390/ijms25095018 - 4 May 2024
Cited by 1 | Viewed by 2066
Abstract
NanoLuc-mediated bioluminescence resonance energy transfer (NanoBRET) has gained popularity for its ability to homogenously measure ligand binding to G protein-coupled receptors (GPCRs), including the subfamily of chemokine receptors. These receptors, such as ACKR3, CXCR4, CXCR3, play a crucial role in the regulation of [...] Read more.
NanoLuc-mediated bioluminescence resonance energy transfer (NanoBRET) has gained popularity for its ability to homogenously measure ligand binding to G protein-coupled receptors (GPCRs), including the subfamily of chemokine receptors. These receptors, such as ACKR3, CXCR4, CXCR3, play a crucial role in the regulation of the immune system, are associated with inflammatory diseases and cancer, and are seen as promising drug targets. The aim of this study was to optimize NanoBRET-based ligand binding to NLuc-ACKR3 and NLuc-CXCR4 using different fluorescently labeled chemokine CXCL12 analogs and their use in a multiplex NanoBRET binding assay of two chemokine receptors at the same time. The four fluorescent CXCL12 analogs (CXCL12-AZD488, -AZD546, -AZD594, -AZD647) showed high-affinity saturable binding to both NLuc-ACKR3 and NLuc-CXCR4, with relatively low levels of non-specific binding. Additionally, the binding of all AZDye-labeled CXCL12s to Nluc receptors was inhibited by pharmacologically relevant unlabeled chemokines and small molecules. The NanoBRET binding assay for CXCL10-AZD488 binding to Nluc-CXCR3 was also successfully established and successfully employed for the simultaneous measurement of the binding of unlabeled small molecules to NLuc-CXCR3 and NLuc-CXCR4. In conclusion, multiplexing the NanoBRET-based competition binding assay is a promising tool for testing unlabeled (small) molecules against multiple GPCRs simultaneously. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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Review

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10 pages, 2518 KiB  
Review
Emerging Synthetic Bioluminescent Reactions for Non-Invasive Imaging of Freely Moving Animals
by Takahiro Kuchimaru
Int. J. Mol. Sci. 2024, 25(13), 7338; https://doi.org/10.3390/ijms25137338 - 4 Jul 2024
Cited by 1 | Viewed by 1910
Abstract
Bioluminescence imaging (BLI) is an indispensable technique for visualizing the dynamics of diverse biological processes in mammalian animal models, including cancer, viral infections, and immune responses. However, a critical scientific challenge remains: non-invasively visualizing homeostatic and disease mechanisms in freely moving animals to [...] Read more.
Bioluminescence imaging (BLI) is an indispensable technique for visualizing the dynamics of diverse biological processes in mammalian animal models, including cancer, viral infections, and immune responses. However, a critical scientific challenge remains: non-invasively visualizing homeostatic and disease mechanisms in freely moving animals to understand the molecular basis of exercises, social behavior, and other phenomena. Classical BLI relies on prolonged camera exposure to accumulate the limited number of photons that traveled from deep tissues in anesthetized or constrained animals. Recent advancements in synthetic bioluminescence reactions, utilizing artificial luciferin–luciferase pairs, have considerably increased the number of detectable photons from deep tissues, facilitating high-speed BLI to capture moving objects. In this review, I provide an overview of emerging synthetic bioluminescence reactions that enable the non-invasive imaging of freely moving animals. This approach holds the potential to uncover unique physiological processes that are inaccessible with current methodologies. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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Other

Jump to: Research, Review

8 pages, 1239 KiB  
Brief Report
In Vivo Luciferin–Luciferase Reaction in Micro-Mini Pigs Using Xenogeneic Rat Bone Marrow Transplantation
by Tomoyuki Abe, Kazuhiro Endo, Yutaka Hanazono and Eiji Kobayashi
Int. J. Mol. Sci. 2024, 25(16), 8609; https://doi.org/10.3390/ijms25168609 - 7 Aug 2024
Cited by 1 | Viewed by 1264
Abstract
Luminescent technology based on the luciferin–luciferase reaction has been extensively employed across various disciplines as a quantitative imaging modality. Owing to its non-invasive imaging capacity, it has evolved as a valuable in vivo bioimaging tool, particularly in small animal models in fields such [...] Read more.
Luminescent technology based on the luciferin–luciferase reaction has been extensively employed across various disciplines as a quantitative imaging modality. Owing to its non-invasive imaging capacity, it has evolved as a valuable in vivo bioimaging tool, particularly in small animal models in fields such as gene and cell therapies. We have previously successfully generated rats with a systemic expression of the luciferase gene at the Rosa26 locus. In this study, we transplanted bone marrow from these rats into micro-mini pigs and used in vivo imaging to non-invasively analyze the dynamics of the transplanted cells. In addition, we established that the rat-to-pig transplantation system is a discordant system, similar to the pig-to-human transplantation system. Thus, rat-to-pig transplantation may provide a clinically appropriate large animal model for pig-to-human xenotransplantation. Full article
(This article belongs to the Special Issue Advances in Luciferase)
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