Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting guide RNA prevents wet age-related macular degeneration in mice.
Animals
CRISPR-Associated Protein 9
/ genetics
CRISPR-Cas Systems
Disease Models, Animal
Gene Editing
/ methods
Genetic Vectors
/ physiology
HEK293 Cells
Humans
Lentivirus
/ physiology
Macular Degeneration
/ genetics
Mice
Mice, Inbred C57BL
RNA, Messenger
/ genetics
Vascular Endothelial Growth Factor A
/ genetics
Journal
Nature biomedical engineering
ISSN: 2157-846X
Titre abrégé: Nat Biomed Eng
Pays: England
ID NLM: 101696896
Informations de publication
Date de publication:
02 2021
02 2021
Historique:
received:
03
12
2019
accepted:
03
11
2020
pubmed:
6
1
2021
medline:
13
3
2021
entrez:
5
1
2021
Statut:
ppublish
Résumé
Therapeutic genome editing requires effective and targeted delivery methods. The delivery of Cas9 mRNA using adeno-associated viruses has led to potent in vivo therapeutic efficacy, but can cause sustained Cas9 expression, anti-Cas9 immune responses and off-target edits. Lentiviral vectors have been engineered to deliver nucleases that are expressed transiently, but in vivo evidence of their biomedical efficacy is lacking. Here, we show that the lentiviral codelivery of Streptococcus pyogenes Cas9 mRNA and expression cassettes that encode a guide RNA that targets vascular endothelial growth factor A (Vegfa) is efficacious in a mouse model of wet age-related macular degeneration induced by Vegfa. A single subretinal injection of engineered lentiviruses knocked out 44% of Vegfa in retinal pigment epithelium and reduced the area of choroidal neovascularization by 63% without inducing off-target edits or anti-Cas9 immune responses. Engineered lentiviruses for the transient expression of nucleases may form the basis of new treatments for retinal neovascular diseases.
Identifiants
pubmed: 33398131
doi: 10.1038/s41551-020-00656-y
pii: 10.1038/s41551-020-00656-y
doi:
Substances chimiques
RNA, Messenger
0
Vascular Endothelial Growth Factor A
0
vascular endothelial growth factor A, mouse
0
CRISPR-Associated Protein 9
EC 3.1.-
Banques de données
figshare
['10.6084/m9.figshare.12611819']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
144-156Commentaires et corrections
Type : CommentIn
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