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Protein Delivery: If Your GFP (or Other Small Protein) Is in the Cytosol, It Will Also Be in the Nucleus

  • David C. Luther
    David C. Luther
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
  • Taewon Jeon
    Taewon Jeon
    Molecular and Cellular Biology Graduate Program, University of Massachusetts, 230 Stockbridge Road, Amherst, Massachusetts 01003, United States
    More by Taewon Jeon
  • Ritabrita Goswami
    Ritabrita Goswami
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
  • Harini Nagaraj
    Harini Nagaraj
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
  • Dongkap Kim
    Dongkap Kim
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
    Center for Biomaterials, Biomedical Research Institute, Korea Institute of Science and Technology (KIST), Seoul, 02792, Republic of Korea
    Department of Chemistry, Hanyang University, Seoul 04763, Republic of Korea
    More by Dongkap Kim
  • Yi-Wei Lee
    Yi-Wei Lee
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
    More by Yi-Wei Lee
  • , and 
  • Vincent M. Rotello*
    Vincent M. Rotello
    Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States
    *Email: [email protected]
Cite this: Bioconjugate Chem. 2021, 32, 5, 891–896
Publication Date (Web):April 19, 2021
https://doi.org/10.1021/acs.bioconjchem.1c00103
Copyright © 2021 American Chemical Society

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    Abstract

    Abstract Image

    Intracellular protein delivery is a transformative tool for biologics research and medicine. Delivery into the cytosol allows proteins to diffuse throughout the cell and access subcellular organelles. Inefficient delivery caused by endosomal entrapment is often misidentified as cytosolic delivery. This inaccuracy muddles what should be a key checkpoint in assessing delivery efficiency. Green fluorescent protein (GFP) is a robust cargo small enough to passively diffuse from the cytosol into the nucleus. Fluorescence of GFP in the nucleus is a direct readout for cytosolic access and effective delivery. Here, we highlight recent examples from the literature for the accurate assessment of cytosolic protein delivery using GFP fluorescence in the cytosol and nucleus.

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    Cited By

    This article is cited by 18 publications.

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