Premium
Silicon‐Nanotube‐Mediated Intracellular Delivery Enables Ex Vivo Gene Editing
Author(s) -
Chen Yaping,
Aslanoglou Stella,
Murayama Takahide,
Gervinskas Gediminas,
Fitzgerald Laura I.,
Sriram Sharath,
Tian Jie,
Johnston Angus P. R.,
Morikawa Yasuhiro,
Suu Koukou,
Elnathan Roey,
Voelcker Nicolas H.
Publication year - 2020
Publication title -
advanced materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 10.707
H-Index - 527
eISSN - 1521-4095
pISSN - 0935-9648
DOI - 10.1002/adma.202000036
Subject(s) - microbiology and biotechnology , endocytic cycle , intracellular , endocytosis , ex vivo , gene delivery , materials science , nanotechnology , genome editing , biology , biophysics , cell , transfection , crispr , gene , biochemistry , in vitro
Engineered nano–bio cellular interfaces driven by vertical nanostructured materials are set to spur transformative progress in modulating cellular processes and interrogations. In particular, the intracellular delivery—a core concept in fundamental and translational biomedical research—holds great promise for developing novel cell therapies based on gene modification. This study demonstrates the development of a mechanotransfection platform comprising vertically aligned silicon nanotube (VA‐SiNT) arrays for ex vivo gene editing. The internal hollow structure of SiNTs allows effective loading of various biomolecule cargoes; and SiNTs mediate delivery of those cargoes into GPE86 mouse embryonic fibroblasts without compromising their viability. Focused ion beam scanning electron microscopy (FIB‐SEM) and confocal microscopy results demonstrate localized membrane invaginations and accumulation of caveolin‐1 at the cell–NT interface, suggesting the presence of endocytic pits. Small‐molecule inhibition of endocytosis suggests that active endocytic process plays a role in the intracellular delivery of cargo from SiNTs. SiNT‐mediated siRNA intracellular delivery shows the capacity to reduce expression levels of F‐actin binding protein (Triobp) and alter the cellular morphology of GPE86. Finally, the successful delivery of Cas9 ribonucleoprotein (RNP) to specifically target mouse Hprt gene is achieved. This NT‐enhanced molecular delivery platform has strong potential to support gene editing technologies.