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

  title={Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting guide RNA prevents wet age-related macular degeneration in mice},
  author={Sikai Ling and Shiqi Yang and Xinde Hu and Di Yin and Yao Dai and Xiaoqin Qian and Dawei Wang and Xiaoyong Pan and Jiaxu Hong and Xiaodong Sun and Hui Yang and Soren Riis Paludan and Yujia Cai},
  journal={Nature Biomedical Engineering},
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… Expand
4 Citations
CRISPR Cas9 based genome editing in inherited retinal dystrophies
The CRISPR-Cas9 system has revolutionized genome editing, and opened avenues in drug discovery, and it is important to understand the role of this system along with its applicability in the field of ophthalmology. Expand
In vivo delivery of CRISPR-Cas9 therapeutics: Progress and challenges
  • Matthew Behr, Jing Zhou, Bing Xu, Hongwei Zhang
  • Medicine
  • Acta pharmaceutica Sinica. B
  • 2021
The most recent progress of in vivo delivery of CRISPR-Cas9 systems is discussed, innovative viral and non-viral delivery technologies are highlighted, outstanding delivery challenges are emphasized, and the most updated perspectives are provided. Expand
Ocular Therapeutics and Molecular Delivery Strategies for Neovascular Age-Related Macular Degeneration (nAMD)
The present review aims to highlight the status of pre-clinical and clinical studies of neovascular AMD treatment modalities including Anti-VEGF therapy, upcoming bispecific antibodies, small molecules, port delivery systems, photodynamic therapy, radiation therapy, gene therapy, cell therapy, and combination therapies. Expand
High-purity production and precise editing of DNA base editing ribonucleoproteins
  • Hyeon-Ki Jang, D. Jo, +7 authors Sangsu Bae
  • Medicine
  • Science advances
  • 2021
Cytosine and adenine base editor ribonucleoproteins show precise base editing with reduced DNA and RNA off-target effects.


In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells
The capacity of lentivirus-delivered SpCas9 and sgRNAs as a developing therapeutic path in the treatment of ocular diseases, including age-related macular degeneration is demonstrated. Expand
Adenovirus-Mediated Somatic Genome Editing of Pten by CRISPR/Cas9 in Mouse Liver in Spite of Cas9-Specific Immune Responses
It is found that the Ad vector mediated efficient Pten gene editing even in the presence of typical Ad vector-associated immunotoxicity in the liver, consistent with the phenotypes following Cre-loxP-induced Pten deficiency in mouse liver. Expand
Genome surgery using Cas9 ribonucleoproteins for the treatment of age-related macular degeneration.
The results demonstrate that in vivo genome editing with Cas9 RNPs has the potential for the local treatment for nongenetic degenerative diseases, expanding the scope of RNA-guided genome surgery to a new dimension. Expand
Inhibition of choroidal neovascularization via brief subretinal exposure to a newly developed lentiviral vector pseudotyped with Sendai viral envelope proteins.
It is demonstrated that brief subretinal administration of SeV-F/HN-SIV vectors may facilitate safe and efficient retinal gene transfer, and the therapeutic potential of PEDF with a higher safety profile for treating CNV in AMD patients is suggested. Expand
Integrase-Deficient Lentiviral Vector as an All-in-One Platform for Highly Efficient CRISPR/Cas9-Mediated Gene Editing
An all-in-one vector cassette is developed with increased production efficacy and demonstrated that CRISPR/Cas9 delivered by the improved IDLV vectors can mediate rapid and robust gene editing in human embryonic kidney cells and post-mitotic brain neurons in vivo, via transient expression and with higher gene-targeting specificity than the corresponding integrase-competent vectors. Expand
Highly efficient in vitro and in vivo delivery of functional RNAs using new versatile MS2-chimeric retrovirus-like particles
The novel chimeric MS2-lentiviral particles are a versatile tool for a wide range of applications including cellular-programming or genome-editing and activate an osteoblast differentiation pathway by delivering RUNX2- or DLX5-mRNA into primary human bone-marrow mesenchymal-stem cells. Expand
Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
This work has developed a system able to package up to 100 copies of Staphylococcus aureus Cas9 (SaCas9) mRNA in each lentivirus-like bionanoparticle (LVLP) and achieves highly efficient genome editing in the presence of guide RNA. Expand
Lentivirus pre-packed with Cas9 protein for safer gene editing
Results show that sgRNA/Cas9P LV can be used as a safer approach for human gene therapy applications, and is able to disrupt HIV provirus in the J-LAT model of viral latency. Expand
A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing
A self-deleting AAV-CRISPR system that introduces insertion and deletion mutations into AAV episomes that dramatically reduces the level of Staphylococcus aureus Cas9 protein, while achieving high rates of on-target editing in the liver. Expand
Targeted deletion of Vegfa in adult mice induces vision loss.
It is shown that conditionally knocking out Vegfa in adult mouse retinal pigmented epithelial (RPE) cells, which regulate retinal homeostasis, rapidly leads to vision loss and ablation of the choriocapillaris, the major blood supply for the outer retina and photoreceptor cells. Expand