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A molecular sensor to quantify the localization of proteins, DNA and nanoparticles in cells

Author

Listed:
  • Laura I. FitzGerald

    (Monash University
    Monash University)

  • Luigi Aurelio

    (Monash University)

  • Moore Chen

    (Monash University)

  • Daniel Yuen

    (Monash University)

  • Joshua J. Rennick

    (Monash University
    Monash University)

  • Bim Graham

    (Monash University)

  • Angus P. R. Johnston

    (Monash University
    Monash University)

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.

Suggested Citation

  • Laura I. FitzGerald & Luigi Aurelio & Moore Chen & Daniel Yuen & Joshua J. Rennick & Bim Graham & Angus P. R. Johnston, 2020. "A molecular sensor to quantify the localization of proteins, DNA and nanoparticles in cells," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-18082-8
    DOI: 10.1038/s41467-020-18082-8
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    Cited by:

    1. Hampus Hedlund & Hampus Rietz & Johanna M. Johansson & Hanna C. Eriksson & Wahed Zedan & Linfeng Huang & Jonas Wallin & Anders Wittrup, 2023. "Single-cell quantification and dose-response of cytosolic siRNA delivery," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    2. Joshua J. Rennick & Cameron J. Nowell & Colin W. Pouton & Angus P. R. Johnston, 2022. "Resolving subcellular pH with a quantitative fluorescent lifetime biosensor," Nature Communications, Nature, vol. 13(1), pages 1-13, December.

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