Silicon-Nanotube-Mediated Intracellular Delivery Enables Ex Vivo Gene Editing.
Cas9 RNP
gene editing
intracellular delivery
siRNA knockdown
silicon nanotubes
Journal
Advanced materials (Deerfield Beach, Fla.)
ISSN: 1521-4095
Titre abrégé: Adv Mater
Pays: Germany
ID NLM: 9885358
Informations de publication
Date de publication:
Jun 2020
Jun 2020
Historique:
received:
02
01
2020
revised:
16
03
2020
accepted:
22
03
2020
pubmed:
8
5
2020
medline:
13
4
2021
entrez:
8
5
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32378244
doi: 10.1002/adma.202000036
doi:
Substances chimiques
Caveolin 1
0
RNA, Small Interfering
0
Silicon
Z4152N8IUI
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2000036Subventions
Organisme : Australian Government
ID : DE170100021
Organisme : CSIRO
Organisme : CSIRO Research Office for a Science Leader Fellowship
Informations de copyright
© 2020 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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