Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting guide RNA prevents wet age-related macular degeneration in mice
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...
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Veröffentlicht in: | Nature biomedical engineering 2021-02, Vol.5 (2), p.144-156 |
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Sprache: | eng |
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Zusammenfassung: | 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.
Lentiviruses co-packaging Sp
Cas9
mRNA and an expression cassette encoding for a guide RNA targeting the
Vegfa
gene prevent mice from developing wet age-related macular degeneration induced by
Vegfa
. [Summary amended to correct the description of the packaged cargo.] |
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ISSN: | 2157-846X 2157-846X |
DOI: | 10.1038/s41551-020-00656-y |