Open AccessDirect Cytosolic Delivery of Proteins through Coengineering of Proteins and Polymeric Delivery Vehicles
Author(s) -
Yi-Wei Lee,
David C. Luther,
Ritabrita Goswami,
Taewon Jeon,
Vincent P. Clark,
James Elia,
S. Gopalakrishnan,
Vincent M. Rotello
Publication year - 2020
Publication title -
journal of the american chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.115
H-Index - 612
eISSN - 1520-5126
pISSN - 0002-7863
DOI - 10.1021/jacs.9b12759
Subject(s) - cytosol , chemistry , nanocarriers , endosome , organelle , biophysics , drug delivery , flow cytometry , gene delivery , confocal microscopy , nanotechnology , microbiology and biotechnology , biochemistry , cell , biology , materials science , enzyme , genetic enhancement , organic chemistry , gene
Nanocarrier-mediated protein delivery is a promising strategy for fundamental research and therapeutic applications. However, the efficacy of the current platforms for delivery into cells is limited by endosomal entrapment of delivered protein cargo with concomitantly inefficient access to the cytosol and other organelles, including the nucleus. We report here a robust, versatile polymeric-protein nanocomposite (PPNC) platform capable of efficient (≥90%) delivery of proteins to the cytosol. We synthesized a library of guanidinium-functionalized poly(oxanorborneneimide) (PONI) homopolymers with varying molecular weights to stabilize and deliver engineered proteins featuring terminal oligoglutamate "E-tags". The polymers were screened for cytosolic delivery efficiency using imaging flow cytometry with cytosolic delivery validated using confocal microscopy and activity of the delivered proteins demonstrated through functional assays. These studies indicate that the PPNC platform provides highly effective and tunable cytosolic delivery over a wide range of formulations, making them robust agents for therapeutic protein delivery.