Goldman C, Kareva T, Sarrafha L, Schuldt BR, Sahasrabudhe A, Ahfeldt T, Blanchard JW (2025) Genetically encoded and modular subcellular organelle probes reveal dysfunction in lysosomes and mitochondria driven by PRKN knockout. iScience 28(7). doi: 10.1016/j.isci.2025.112816
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: May 16, 2025
Last Modified: May 29, 2025
Collection Integer ID: 218391
Keywords: ASAPCRN, modular subcellular organelle probe, quantitative visualization of mitochondria, critical insight into the subcellular mechanism, oxidative stress of specific organelle, mitochondria, differentiated astrocyte, knockout model of parkinson, subcellular mechanism, lysosome, mitochondrial transport, mitochondrial dysfunction, organelle, specific organelle, human pluripotent stem cell prkn, parkinson, underlying parkinson, including lysosomal, modular toolbox of fluorescent marker, human cell, cell, gem, oxidative stress, disease in human cell, cellular model, lysosomal distribution, neuron, diversity of cellular model, imaging tool
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-024297
NASA
Grant ID: 80ARC022CA004
NIH/NINDS
Grant ID: R01NS114239
NIH/NINDS
Grant ID: UH3NS115064
NIH/NIA
Grant ID: T32AG04968
NIH/NINDS
Grant ID: F31NS13090
New York State Department of Health
Grant ID: NYSTEM-C32561GG
Abstract
Cellular processes including lysosomal and mitochondrial dysfunction are implicated in the development of many diseases. Quantitative visualization of mitochondria and lysosomes is crucial to understand how these organelles are dysregulated during disease. To address a gap in live-imaging tools, we developed GEM-SCOPe (Genetically Encoded and Modular SubCellular Organelle Probes), a modular toolbox of fluorescent markers designed to inform on localization, distribution, turnover, and oxidative stress of specific organelles. We expressed GEM-SCOPe in differentiated astrocytes and neurons from a human pluripotent stem cell PRKN-knockout model of Parkinson’s disease and identified disease-associated changes in proliferation, lysosomal distribution, mitochondrial transport and turnover, and reactive oxygen species. We demonstrate GEM-SCOPe is a powerful panel that provide critical insight into the subcellular mechanisms underlying Parkinson’s disease in human cells. GEM-SCOPe can be expanded upon and applied to a diversity of cellular models to glean an understanding of the mechanisms that promote disease onset and progression.
