Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting guide RNA prevents wet age-related macular degeneration in mice

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Sikai Ling, Shanghai Jiao Tong University
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  • Shiqi Yang, Shanghai Jiao Tong University
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  • Xinde Hu, CAS - Shanghai Institute for Biological Sciences
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  • Di Yin, Shanghai Jiao Tong University
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  • Yao Dai, Shanghai Jiao Tong University
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  • Xiaoqing Qian, Shanghai Jiao Tong University
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  • Dawei Wang, Shanghai Jiao Tong University
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  • Xiaoyong Pan, Shanghai Jiao Tong University
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  • Jiaxu Hong, Fudan University
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  • Xiaodong Sun, Shanghai Jiao Tong University
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  • Hui Yang, CAS - Shanghai Institute for Biological Sciences
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  • Soren Riis Paludan
  • Yujia Cai, Shanghai Jiao Tong University

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 pyogenesCas9 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.

Original languageEnglish
JournalNature Biomedical Engineering
Pages (from-to)144-156
Number of pages13
Publication statusPublished - Feb 2021

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