Genetically Encoded, pH-Sensitive mTFP1 Biosensor for Probing Lysosomal pH
Marcus Y. Chin, Anand Patwardhan, Kean-Hooi Ang, Austin L. Wang, Carolina Alquézar, Mackenzie Welch, Phi T. Nguyen, Michael Grabe, Anna V. Molofsky, Michelle R. Arkin, Aimee W. Kao
Abstract
of 4.3. Ratiometric quantification was enabled with cytosolically oriented mCherry using high-content quantitative imaging. We expressed FIRE-pHLy in several cellular models and quantified the alkalinizing response to bafilomycin A1, a specific V-ATPase inhibitor. In summary, we have engineered FIRE-pHLy, a specific, robust, and versatile lysosomal pH biosensor, that has broad applications for investigating pH dynamics in aging- and lysosome-related diseases, as well as in lysosome-based drug discovery.
Topics & Concepts
BafilomycinLysosomeBiosensorChemistryFluorescenceBiochemistrymCherryMannose 6-phosphate receptorGreen fluorescent proteinBiophysicsBiologyEnzymeAutophagyApoptosisPhysicsGeneQuantum mechanicsAdvanced Fluorescence Microscopy TechniquesCellular transport and secretionCalcium signaling and nucleotide metabolism