Projects per year
Abstract
Intracellular trafficking governs receptor signaling, pathogenesis, immune responses and fate of nanomedicines. These processes are typically tracked by observing colocalization of fluorescent markers using confocal microscopy. However, this method is low throughput, limited by the resolution of microscopy, and can miss fleeting interactions. To address this, we developed a localization sensor composed of a quenched SNAP-tag substrate (SNAPSwitch) that can be conjugated to biomolecules using click chemistry. SNAPSwitch enables quantitative detection of trafficking to locations of interest within live cells using flow cytometry. Using SNAPSwitch, we followed the trafficking of DNA complexes from endosomes into the cytosol and nucleus. We show that antibodies against the transferrin or hyaluronan receptor are initially sorted into different compartments following endocytosis. In addition, we can resolve which side of the cellular membrane material was located. These results demonstrate SNAPSwitch is a high-throughput and broadly applicable tool to quantitatively track localization of materials in cells.
Original language | English |
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Article number | 4482 |
Number of pages | 13 |
Journal | Nature Communications |
Volume | 11 |
Issue number | 1 |
DOIs | |
Publication status | Published - 1 Dec 2020 |
Projects
- 1 Finished
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ARC Centre of Excellence in Convergent Bio-Nano Science and Technology
Davis, T. (Primary Chief Investigator (PCI)), Boyd, B. (Chief Investigator (CI)), Bunnett, N. (Chief Investigator (CI)), Porter, C. (Chief Investigator (CI)), Caruso, F. (Chief Investigator (CI)), Kent, S. (Chief Investigator (CI)), Thordarson, P. (Chief Investigator (CI)), Kearnes, M. (Chief Investigator (CI)), Gooding, J. (Chief Investigator (CI)), Kavallaris, M. (Chief Investigator (CI)), Thurecht, K. (Chief Investigator (CI)), Whittaker, A. K. (Chief Investigator (CI)), Parton, R. (Chief Investigator (CI)), Corrie, S. R. (Chief Investigator (CI)), Johnston, A. (Chief Investigator (CI)), McGhee, J. (Chief Investigator (CI)), Greguric, I. D. (Partner Investigator (PI)), Stevens, M. M. (Partner Investigator (PI)), Lewis, J. S. (Partner Investigator (PI)), Lee, D. S. (Partner Investigator (PI)), Alexander, C. (Partner Investigator (PI)), Dawson, K. (Partner Investigator (PI)), Hawker, C. (Partner Investigator (PI)), Haddleton, D. (Partner Investigator (PI)), Thierry, B. (Chief Investigator (CI)), Prestidge, C. A. (Chief Investigator (CI)), Meyer, A. (Project Manager), Jones-Jayasinghe, N. (Project Manager), Voelcker, N. (Chief Investigator (CI)), Nann, T. (Chief Investigator (CI)) & McLean, K. (Partner Investigator (PI))
Australian Research Council (ARC), Monash University, University of Melbourne, University of New South Wales (UNSW), University of Queensland , University of South Australia, Monash University – Internal Faculty Contribution, University of Wisconsin Madison, Memorial Sloan Kettering Cancer Center, University of California System, University College Dublin, Imperial College London, University of Warwick, Sungkyunkwan University, Australian Nuclear Science and Technology Organisation (ANSTO) , University of Nottingham
30/06/14 → 29/06/21
Project: Research