Humanization and expression of Macrolampis sp2 firefly luciferase for intracelular pH-biosensing

• Grant number: 15/22603-4
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 01, 2016
• End date: June 30, 2016
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Gabriele Verônica de Mello Gabriel
• Supervisor: Yoshihiro Ohmiya
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
• Institution abroad: National Institute of Advanced Industrial Science and Technology, Tsukuba (AIST), Japan
• Associated to the scholarship: 14/04477-9 - Developing of bioluminescent ratiometric biosensors for pH and divalent metals based on the engineering of the pH-sensing moiety of firefly luciferases, BP.DR

Abstract

Bioluminescence, the emission of visible light by living organisms, is widely used in biosensors. Firefly luciferases genes are among the most used reporter genes in bioluminescent biosensors. Firefly luciferases are pH-sensitive displaying a large red shift at acid pH, a property that has been considered a useless for most applications. However, currently intracellular pH-biosensors are on demand, some of them are fluorescent and ratiometric. Thus we thought to use firefly luciferase pH-sensitivity to advantage of measuring intracellular pH ratiometrically. We already showed a relationship between pH and ratio of intensities of bioluminescence spectra in the green and red region that could be used to ratiometrically estimate intracellular pH in bacteria. The goal of this project is to use Macrolampis sp2 firefly luciferase as intracellular pH biosensor for mammalian cells. For this purpose, we aim to humanize Macrolampis sp2 luciferase, express it in mammalian cells and to investigate intracellular pH variations ratiometrically.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE MELLO GABRIEL, GABRIELE VERONICA; YASUNO, RIE; MITANI, YASUO; OHMIYA, YOSHIHIRO; VIVIANI, VADIM RAVARA. Novel application of Macrolampis sp2 firefly luciferase for intracellular pH-biosensing in mammalian cells. PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, v. 18, n. 5, p. 1212-1217, MAY 1 2019. (10/05426-8, 14/04477-9, 16/15946-5, 11/23961-0, 15/22603-4)