Quantitative Bioluminescent Imaging of AHR Activity Using a Thermostable Variant of the Fungal Luciferase nnLuz
9 affiliations
- 1Epigenetic Memory Group, MRC Laboratory of Medical Sciences, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0HS, UK
- 2Planta LLC, Skolkovo Innovation Center, Bolshoy Boulevard 42, Building 1, 335, Moscow 121205, Russia
- 3Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia
- 4Whole Animal Physiology and Imaging, MRC Laboratory of Medical Sciences, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0HS, UK
- 5Transgenics & Embryonic Stem Cell Facility, MRC Laboratory of Medical Sciences, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0HS, UK
- 6Synthetic Biology Group, MRC Laboratory of Medical Sciences, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0HS, UK
- 7Lymphocyte Development Group, MRC Laboratory of Medical Sciences, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0HS, UK
- 8Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK
- 9Institute of Clinical Sciences, Faculty of Medicine and Imperial College Centre for Engineering Biology, Imperial College London, London W12 0HS, UK
Abstract
Bioluminescent reporters are important tools in biomedical research. Luciferase enzymes, expressed as part of transgenes or inserted into endogenous genomes, can be used to visualise molecular events with minimally invasive experiments. However, the reaction requires addition of the substrate, luciferin. Organisms with self-sustained luminescence that both express the luciferase and produce the luciferin would allow continuous, convenient and affordable acquisition of physiological information. The Neonothopanus nambi fungal bioluminescence pathway has been harnessed in this way to produce autoluminescent plants. However, progress into mammalian models has been hindered by temperature sensitivity of the wild-type fungal luciferase (nnLuz). We tested the efficacy of a thermostable variant of nnLuz (nnLuz_v3) in vivo at physiologically relevant levels, by generating a reporter of endogenous Cyp1a1 expression as a measure of aryl hydrocarbon receptor (AHR) activation. Previously published Cyp1a1-Fluc mice allowed a comparison between the novel luciferase nnLuz_v3 and the established firefly luciferase (Fluc). In Cyp1a1-nnLuz_v3 mice, the bioluminescence signal was sufficiently stable to enable well-resolved luminescence images to be captured on standard in vivo imaging equipment and consumer-grade cameras. These results derisk the use of nnLuz_v3 as an in vivo molecular reporter whilst providing a novel tool for AHR drug discovery and research.
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BibTeX
@article{gleneadie2026quantitative,
title = {{Quantitative Bioluminescent Imaging of AHR Activity Using a Thermostable Variant of the Fungal Luciferase nnLuz}},
author = {Hannah J. Gleneadie and Nicolas Veland and Nadezhda M. Markina and Alessandro Sardini and Anastasia A. Fadeeva and Zoe Webster and Feng Gao and Matthias Merkenschlager and Amanda G. Fisher and Karen S. Sarkisyan},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27219586},
url = {https://www.mdpi.com/1422-0067/27/21/9586},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Quantitative Bioluminescent Imaging of AHR Activity Using a Thermostable Variant of the Fungal Luciferase nnLuz
AU - Hannah J. Gleneadie
AU - Nicolas Veland
AU - Nadezhda M. Markina
AU - Alessandro Sardini
AU - Anastasia A. Fadeeva
AU - Zoe Webster
AU - Feng Gao
AU - Matthias Merkenschlager
AU - Amanda G. Fisher
AU - Karen S. Sarkisyan
PY - 2026
DA - 2026/10/27
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219586
UR - https://www.mdpi.com/1422-0067/27/21/9586
AB - Bioluminescent reporters are important tools in biomedical research. Luciferase enzymes, expressed as part of transgenes or inserted into endogenous genomes, can be used to visualise molecular events with minimally invasive experiments. However, the reaction requires addition of the substrate, luciferin. Organisms with self-sustained luminescence that both express the luciferase and produce the luciferin would allow continuous, convenient and affordable acquisition of physiological information. The Neonothopanus nambi fungal bioluminescence pathway has been harnessed in this way to produce autoluminescent plants. However, progress into mammalian models has been hindered by temperature sensitivity of the wild-type fungal luciferase (nnLuz). We tested the efficacy of a thermostable variant of nnLuz (nnLuz_v3) in vivo at physiologically relevant levels, by generating a reporter of endogenous Cyp1a1 expression as a measure of aryl hydrocarbon receptor (AHR) activation. Previously published Cyp1a1-Fluc mice allowed a comparison between the novel luciferase nnLuz_v3 and the established firefly luciferase (Fluc). In Cyp1a1-nnLuz_v3 mice, the bioluminescence signal was sufficiently stable to enable well-resolved luminescence images to be captured on standard in vivo imaging equipment and consumer-grade cameras. These results derisk the use of nnLuz_v3 as an in vivo molecular reporter whilst providing a novel tool for AHR drug discovery and research.
SN - 1422-0067
ER -