Fabrication and use of silicon hollow-needle arrays to achieve tissue nanotransfection in mouse tissue in vivo.
Animals
Cellular Reprogramming Techniques
/ methods
Electroporation
/ methods
Male
Mice
Mice, Inbred C57BL
Microtechnology
/ instrumentation
Nanotechnology
/ instrumentation
Oligonucleotide Array Sequence Analysis
/ instrumentation
Plasmids
/ chemistry
Quality Control
Silicon
/ chemistry
Skin
/ metabolism
Transfection
/ instrumentation
Journal
Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307
Informations de publication
Date de publication:
12 2021
12 2021
Historique:
received:
14
06
2020
accepted:
10
09
2021
pubmed:
28
11
2021
medline:
24
12
2021
entrez:
27
11
2021
Statut:
ppublish
Résumé
Tissue nanotransfection (TNT) is an electromotive gene transfer technology that was developed to achieve tissue reprogramming in vivo. This protocol describes how to fabricate the required hardware, commonly referred to as a TNT chip, and use it for in vivo TNT. Silicon hollow-needle arrays for TNT applications are fabricated in a standardized and reproducible way. In <1 s, these silicon hollow-needle arrays can be used to deliver plasmids to a predetermined specific depth in murine skin in response to pulsed nanoporation. Tissue nanotransfection eliminates the need to use viral vectors, minimizing the risk of genomic integration or cell transformation. The TNT chip fabrication process typically takes 5-6 d, and in vivo TNT takes 30 min. This protocol does not require specific expertise beyond a clean room equipped for basic nanofabrication processes.
Identifiants
pubmed: 34837085
doi: 10.1038/s41596-021-00631-0
pii: 10.1038/s41596-021-00631-0
pmc: PMC9104164
mid: NIHMS1785546
doi:
Substances chimiques
Silicon
Z4152N8IUI
Banques de données
figshare
['10.6084/m9.figshare.16528311']
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Review
Langues
eng
Sous-ensembles de citation
IM
Pagination
5707-5738Subventions
Organisme : NIGMS NIH HHS
ID : K25 GM143572
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK114718
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK128845
Pays : United States
Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer Nature Limited.
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