Open AccessCytosolic Delivery of Functional Proteins In Vitro through Tunable Gigahertz Acoustics
Author(s) -
Shuting Pan,
Taewon Jeon,
David C. Luther,
Xuexin Duan,
Vincent M. Rotello
Publication year - 2020
Publication title -
acs applied materials and interfaces
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.535
H-Index - 228
eISSN - 1944-8252
pISSN - 1944-8244
DOI - 10.1021/acsami.9b21131
Subject(s) - cytosol , materials science , intracellular , biophysics , nanotechnology , gene delivery , microfluidics , membrane permeability , in vitro , membrane , cell culture , microbiology and biotechnology , chemistry , biochemistry , biology , transfection , enzyme , genetics
Intracellular delivery is essential to therapeutic applications such as genome engineering and disease diagnosis. Current methods lack simple, noninvasive strategies and are often hindered by long incubation time or high toxicity. Hydrodynamic approaches offer rapid and controllable delivery of small molecules, but thus far have not been demonstrated for delivering functional proteins. In this work, we developed a robust hydrodynamic approach based on gigahertz (GHz) acoustics to achieve rapid and noninvasive cytosolic delivery of biologically active proteins. With this method, GHz-based acoustic devices trigger oscillations through a liquid medium (acoustic streaming), generating shear stress on the cell membrane and inducing transient nanoporation. This mechanical effect enhances membrane permeability and enables cytosolic access to cationic proteins without disturbing their bioactivity. We evaluated the versatility of this approach through the delivery of cationic fluorescent proteins to a range of cell lines, all of which displayed equally efficient delivery speed (≤20 min). Delivery of multiple enzymatically active proteins with functionality related to apoptosis or genetic recombination further demonstrated the relevance of this method.