Cytosolic Delivery of Macromolecules in Live Human Cells Using the Combined Endosomal Escape Activities of a Small Molecule and Cell Penetrating Peptides

Ineffective cellular delivery is a common problem in numerous biological applications. Developing delivery reagents that work robustly in a variety of experimental settings remains a challenge. Herein, we report how peptides derived from the prototypical cell penetrating peptide TAT can be used in c...

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Bibliographic Details
Published in:ACS chemical biology 2019-12, Vol.14 (12), p.2641-2651
Main Authors: Allen, Jason, Najjar, Kristina, Erazo-Oliveras, Alfredo, Kondow-McConaghy, Helena M, Brock, Dakota J, Graham, Kristin, Hager, Elizabeth C, Marschall, Andrea L. J, Dübel, Stefan, Juliano, Rudolph L, Pellois, Jean-Philippe
Format: Article
Language:English
Subjects:
Cell-Penetrating Peptides - metabolism
Cytosol - drug effects
Cytosol - metabolism
Endosomes - drug effects
Endosomes - metabolism
Humans
Macromolecular Substances - metabolism
Pyrazines - pharmacology
Pyridines - pharmacology
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Summary:Ineffective cellular delivery is a common problem in numerous biological applications. Developing delivery reagents that work robustly in a variety of experimental settings remains a challenge. Herein, we report how peptides derived from the prototypical cell penetrating peptide TAT can be used in combination with a small molecule, UNC7938, to deliver macromolecules into the cytosol of cells by a simple co-incubation protocol. We establish successful delivery of peptides, DNA plasmids, and a single-chain variable fragment antibody. We also demonstrate that delivery works in hard-to-transfect mammalian cells and under conditions typically inhibitory to cell-penetrating peptides. Mechanistically, UNC7938 destabilizes the membrane of endosomes. This, in turn, enhances the endosome-leakage activity of cell-penetrating peptides and facilitates the endosomal escape of macromolecules initially internalized by mammalian cells via endocytosis. This combined selective membrane-destabilization represents a new chemical space for delivery tools and provides a novel solution to the problem of endosomal entrapment that often limits the effectiveness of reagent-based delivery approaches.
ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.9b00585