Use of a blue-shifted luciferase for intracellular pH analysis and temperature sensing in mitochondria

• Grant number: 23/02482-4
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: August 23, 2023
• End date: August 22, 2024
• Field of knowledge: Engineering - Biomedical Engineering - Bioengineering
• Principal Investigator: Eliana Aparecida de Rezende Duek
• Grantee: Vanessa Rezende Bevilaqua
• Supervisor: Aldo Roda
• Host Institution: Faculdade de Ciências Médicas e da Saúde (FCMS). Pontifícia Universidade Católica de São Paulo (PUC-SP). Sorocaba , SP, Brazil
• Institution abroad: Università di Bologna, Italy
• Associated to the scholarship: 20/07649-6 - Application of bioluminescence for cell tracking and inflammatory processes: in vitro and in vivo study, BP.PD

Abstract

The firefly luciferin-luciferase system has been used in an enormous variety of bioanalytical applications. Firefly luciferase genes have been widely used as reporters in gene expression studies and for cell tracking in biological and pathological processes, including cell proliferation studies, cytotoxicity assays, and metastasis in model animals, emerging as a novel technology replacing animal testing by cell assays, and helping the pharmaceutical industry to find new therapeutic drugs. Among the emerging technologies, firefly luciferases are being used as bioluminescent reporters to track pathogenic bacteria and viral infections and as intracellular pH-sensors. The estimation of intracellular pH is essential for cell homeostasis, cellular stress and intoxication. Intracellular and organelle pH variations are often associated with changes in the cellular cycle, such as cell division and apoptosis, and stress indicating pathologies such as inflammation, allergy and cancer. Recent studies showed evidences that temperature of respiring mitochondria increases considerably to nearly 50°C. However, these studies still lack support using other methodologies. The use of temperature-sensitive luciferases could be an interesting approach. Furthermore one of the interesting models to investigate intracellular pH changes are macrophages during their differentiation. AmyLuc was developed to work in mammalian cells during this pos doctoral research. This luciferase is more blue-shifted, less temperature sensitive, with properties which are desirable for intracellular pH-indication at higher temperatures in mammalian cells, as well as showing potential to be used as intracellular temperature sensor. Therefore, it would be interesting to avail the suitability this luciferase for intracellular pH biosensing in different cells, and also to prospect its potential utility as intracellular organelle temperature sensor. (AU)