Open AccessCytosolic Delivery of Macromolecules in Live Human Cells Using the Combined Endosomal Escape Activities of a Small Molecule and Cell Penetrating Peptides
Author(s) -
Jason K. Allen,
Kristijjar,
Alfredo ErazoOliveras,
Helena M Kondow-McConaghy,
Dakota J. Brock,
Kristin Graham,
Elizabeth Hager,
Andrea L. J. Marschall,
Stefan Dübel,
Rudolph L. Juliano,
JeanPhilippe Pellois
Publication year - 2019
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.9b00585
Subject(s) - endosome , endocytosis , cytosol , cell penetrating peptide , microbiology and biotechnology , transfection , macromolecule , internalization , cell , drug delivery , peptide , cell membrane , endocytic cycle , biophysics , chemistry , biology , biochemistry , enzyme , organic chemistry , gene
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.