Prime Editing of Vascular Endothelial Growth Factor Receptor 2 Attenuates Angiogenesis In Vitro
Authors:
- 1 August 2024
- journal article
- research article
- Published by SAGE Publications in The CRISPR Journal
- Vol. 7 (4) , 188-196
Abstract
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 .Keywords
This publication has 49 references indexed in Scilit:
- Phage-assisted evolution and protein engineering yield compact, efficient prime editorsCell, 2023
- Therapeutic in vivo delivery of gene editing agentsCell, 2022
- Enhanced prime editing systems by manipulating cellular determinants of editing outcomesCell, 2021
- In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9Nature Biotechnology, 2014
- Retinal transduction profiles by high-capacity viral vectorsGene Therapy, 2014
- Alternative treatments to inhibit VEGF in age-related choroidal neovascularisation: 2-year findings of the IVAN randomised controlled trialThe Lancet, 2013
- Axl is essential for VEGF‐A‐dependent activation of PI3K/AktThe EMBO Journal, 2012
- Vascular endothelial growth factor receptor-2: Structure, function, intracellular signalling and therapeutic inhibitionCellular Signalling, 2007
- HIV-1 Integration in the Human Genome Favors Active Genes and Local HotspotsCell, 2002
- Differential inhibition of tumor angiogenesis by tie2 and vascular endothelial growth factor receptor‐2 dominant‐negative receptor mutantsInternational Journal of Cancer, 2000