Nonviral ultrasound-mediated gene delivery in small and large animal models.
Animals
DNA
/ genetics
Disease Models, Animal
Gene Transfer Techniques
/ instrumentation
Genetic Therapy
/ methods
Humans
Mice
Microbubbles
Muscle, Skeletal
/ metabolism
Neoplasms
/ genetics
Neurodegenerative Diseases
/ genetics
Plasmids
/ chemistry
Swine
Swine, Miniature
Ultrasonic Therapy
/ methods
Ultrasonic Waves
Ultrasonography
/ methods
Journal
Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307
Informations de publication
Date de publication:
04 2019
04 2019
Historique:
received:
10
04
2018
accepted:
18
12
2018
pubmed:
26
2
2019
medline:
17
4
2019
entrez:
27
2
2019
Statut:
ppublish
Résumé
Ultrasound-mediated gene delivery (sonoporation) is a minimally invasive, nonviral and clinically translatable method of gene therapy. This method offers a favorable safety profile over that of viral vectors and is less invasive as compared with other physical gene delivery approaches (e.g., electroporation). We have previously used sonoporation to overexpress transgenes in different skeletal tissues in order to induce tissue regeneration. Here, we provide a protocol that could easily be adapted to address various other targets of tissue regeneration or additional applications, such as cancer and neurodegenerative diseases. This protocol describes how to prepare, conduct and optimize ultrasound-mediated gene delivery in both a murine and a porcine animal model. The protocol includes the preparation of a microbubble-DNA mix and in vivo sonoporation under ultrasound imaging. Ultrasound-mediated gene delivery can be accomplished within 10 min. After DNA delivery, animals can be followed to monitor gene expression, protein secretion and other transgene-specific outcomes, including tissue regeneration. This procedure can be accomplished by a competent graduate student or technician with prior experience in ultrasound imaging or in performing in vivo procedures.
Identifiants
pubmed: 30804568
doi: 10.1038/s41596-019-0125-y
pii: 10.1038/s41596-019-0125-y
pmc: PMC6520480
mid: NIHMS1019999
doi:
Substances chimiques
DNA
9007-49-2
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1015-1026Subventions
Organisme : NCI NIH HHS
ID : R01 CA112356
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA211602
Pays : United States
Organisme : NIBIB NIH HHS
ID : R01 EB026094
Pays : United States
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