Luciferases expression in mammalian cells for bioassay purposes and phototoxic effect of luciferins and fluorescent compounds from coleoptera and diptera

• Grant number: 21/03901-5
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): June 01, 2021
• Effective date (End): October 31, 2022
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal researcher: Vadim Viviani
• Grantee: Alessandra Freitas Serain
• Home Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
• Associated research grant: 10/05426-8 - Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications, AP.TEM

Abstract

Bioluminescence, the light emission in living organisms for signaling purposes, has several biotechnological and multidisciplinary applications. Luciferases are the responsible enzymes for catalyzing the light emission through the substrates oxidation (luciferins). In insects, bioluminescence is found in only three orders: Collembola, Diptera and Coleoptera. The bioluminescent system best known under the biochemical aspect is the fireflies system (Coleoptera). In Diptera, bioluminescence is found exclusively in mosquito larvae of the Keroplatidae family, but the structures of luciferins and luciferases are not yet known. The genes that encode firefly luciferases, together with their luciferin, are already widely used as reporter genes in bioimaging of biological and pathological processes, and in biosensors. However, despite the critical moment of the pandemic, which increasingly requires sensitive tests on mammalian cells, in Brazil there are still very few studies that use bioluminescence. Our laboratory has a wide repertoire of cloned and developed beetle luciferases, with different biotechnological applications. Thus, in this project, it is proposed to implement cell culture of bioluminescent mammals that can be used for different purposes. Considering the candidate's previous experience with phototoxicity tests, and the lack of photobiological studies of luciferins, we propose to investigate the phototoxic or photoprotective effect of luciferin and other fluorescent compounds from arthropods in bacteria and mammalian cells, using conventional methods and bioluminescence of ATP to assess cell viability. We also plan to perform the fractionation, purification and characterization of Neoditomiya keroplatin (luciferin) and beetles fluorescent compounds. (AU)