| Autor: |
Ma G; Department of Ophthalmology, The First Affiliated Hospital of Hainan Medical University, Haikou, China.; Department of Ophthalmology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, China., Qi H; Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen, China., Deng H; Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen, China., Dong L; Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen, China., Zhang Q; Department of Ophthalmology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, China., Ma J; Department of Ophthalmology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, China., Yang Y; Ningxia Key Laboratory of Prevention and Control of Common Infectious Diseases, the School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, China., Yan X; Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen, China., Duan Y; Department of Ophthalmology, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Third Hospital of Shanxi Medical University, Tongji Shanxi Hospital, Taiyuan, China., Lei H; Department of Ophthalmology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, China. |
| Abstrakt: |
Vascular endothelial growth factor receptor (VEGFR)-2 is a key switch for angiogenesis, which is observed in various human diseases. In this study, a novel system for advanced prime editing (PE), termed PE6h, is developed, consisting of dual lentiviral vectors: (1) a clustered regularly interspaced palindromic repeat-associated protein 9 (H840A) nickase fused with reverse transcriptase and an enhanced PE guide RNA and (2) a dominant negative (DN) MutL homolog 1 gene with nicking guide RNA. PE6h was used to edit VEGFR2 (c.18315T>A, 50.8%) to generate a premature stop codon (TAG from AAG), resulting in the production of DN-VEGFR2 (787 aa) in human retinal microvascular endothelial cells (HRECs). DN-VEGFR2 impeded VEGF-induced phosphorylation of VEGFR2, Akt, and extracellular signal-regulated kinase-1/2 and tube formation in PE6h-edited HRECs in vitro . Overall, our results highlight the potential of PE6h to inhibit angiogenesis in vivo . |
