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Selection and Engineering of Novel Brighter Bioluminescent Reporter Gene and Color- Tuning Luciferase for pH-Sensing in Mammalian Cells.
Biosensors (Basel)
January 2025
Laboratory of Biochemistry, Molecular Biology and Bioluminescent Systems Technology, Department of Physics, Chemistry and Mathematics, Federal University of Sao Carlos (UFSCAR), Rodovia João Leme dos Santos, km 110, Sorocaba 18052-780, SP, Brazil.
Firefly luciferases have been extensively used for bioanalytical applications, including their use as bioluminescent reporters, biosensors, and for bioimaging biological and pathological processes. Due to their intrinsic pH- sensitivity, in recent years we have demonstrated that firefly luciferases can also be harnessed as color- tuning sensors of intracellular pH. However, it is known that mammalian cells require temperatures higher than 36 °C, which red-shift the bioluminescence spectra of most firefly luciferases, decreasing their activities and the resolution of ratiometric pH analysis.
View Article and Find Full Text PDFPEGylated ATP-Independent Luciferins for Noninvasive High-Sensitivity High-Speed Bioluminescence Imaging.
ACS Chem Biol
January 2025
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, Virginia 22908, United States.
Bioluminescence imaging (BLI) is a powerful, noninvasive imaging method for animal studies. NanoLuc luciferase and its derivatives are attractive bioluminescent reporters recognized for their efficient photon production and ATP independence. However, utilizing them for animal imaging poses notable challenges.
View Article and Find Full Text PDFPilot Screening of TREM1 Inhibitors in Cell-Based Reporter Assays Reflecting TREM1/DAP12 Receptor Assembly and Functionality.
ACS Chem Neurosci
January 2025
Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama 35233, United States.
Proinflammatory TREM1 receptors expressed on myeloid-derived cells have recently been recognized as a new oncogenic target in cancer, including gliomas. They are established chemotherapeutic targets in neurodegenerative Parkinson's and Alzheimer's diseases, and they also contribute to stroke and sepsis severities. TREM1 activation requires the TREM1/DAP12 interaction for receptor clustering and signal transduction coordinated by TREM1 ligands.
View Article and Find Full Text PDFInterference of metal ions on the bioluminescent signal of firefly, Renilla, and NanoLuc luciferases in high-throughput screening assays.
Front Chem
November 2024
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland.
Article Synopsis
- Bioluminescent high-throughput screening (HTS) assays using firefly, Renilla, and NanoLuc luciferases are essential tools in research and drug discovery, but can be affected by various metal ions present in biological and environmental samples.
- A study screened 21 metal ions in three different HTS assays, quantifying their interference with luminescence signals and developing optimized buffer variants for better luciferase performance.
- Findings showed that metal ions significantly impacted bioluminescence signals in a way that correlates with known metal ion-ligand affinities, indicating complex interactions that vary depending on the type of luciferase and assay conditions used.
Bioluminescence Imaging to Study Recombinant Orthopoxvirus Infection in Animal Models.
Methods Mol Biol
December 2024
Poxvirus and Rabies Branch, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Bioluminescent images of viral replication in live animals (in vivo) reveal disease dynamics and effects of medical countermeasures over time. After selecting an appropriate orthopoxvirus animal model for the study, a recombinant virus with the firefly luciferase gene inserted in the genome is used to infect the animals. On the day of bioluminescent imaging, the substrate, D-luciferin, is prepared; animals are sedated and injected with the substrate and IVIS imager is utilized; various bioluminescent images are acquired; then animals recover and are able to continue in the study.
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