AAV5 Delivery of CRISPR/Cas9 Mediates Genome Editing in the Lungs of Young Rhesus Monkeys.
Shun-Qing LiangAndrew W NaviaMichelle RamseierXuntao ZhouMichele MartinezCharles LeeChen ZhouJoae WuJun XieQin SuDan WangTerence R FlotteDaniel G AndersonAlice F TarantalAlex K ShalekGuangping GaoShun-Qing LiangPublished in: Human gene therapy (2024)
Genome editing has the potential to treat genetic diseases in a variety of tissues, including the lung. We have previously developed and validated a dual adeno-associated virus (AAV) CRISPR platform that supports effective editing in the airways of mice. To validate this delivery vehicle in a large animal model, we have shown that intratracheal instillation of CRISPR/Cas9 in AAV5 can edit a housekeeping gene or a disease-related gene in the lungs of young rhesus monkeys. We observed up to 8% editing of angiotensin-converting enzyme 2 (ACE2) in lung lobes after single-dose administration. Single-nuclear RNA sequencing revealed that AAV5 transduces multiple cell types in the caudal lung lobes, including alveolar cells, macrophages, fibroblasts, endothelial cells, and B cells. These results demonstrate that AAV5 is efficient in the delivery of CRISPR/Cas9 in the lung lobes of young rhesus monkeys.
Keyphrases
- crispr cas
- genome editing
- gene therapy
- angiotensin converting enzyme
- single cell
- endothelial cells
- angiotensin ii
- genome wide
- copy number
- middle aged
- induced apoptosis
- gene expression
- cystic fibrosis
- cell therapy
- dna methylation
- high throughput
- oxidative stress
- cell cycle arrest
- type diabetes
- cell death
- extracellular matrix
- bone marrow
- genome wide identification
- cell proliferation
- mesenchymal stem cells
- wild type
- vascular endothelial growth factor