Prime Editing of Vascular Endothelial Growth Factor Receptor 2 Attenuates Angiogenesis In Vitro

被引：1

作者：

Ma, Gaoen ^{[1
,2
]}

Qi, Hui ^{[3
]}

Deng, Hongwei ^{[3
]}

Dong, Lijun ^{[3
]}

Zhang, Qing ^{[2
]}

Ma, Junkai ^{[2
]}

Yang, Yanhui ^{[4
]}

Yan, Xiaohe ^{[3
]}

Duan, Yajian ^{[5
]}

Lei, Hetian ^{[2
]}

机构：

[1] Hainan Med Univ, Affiliated Hosp 1, Dept Ophthalmol, Haikou, Peoples R China

[2] Xinxiang Med Univ, Affiliated Hosp 3, Dept Ophthalmol, 599 Hualan Rd, Xinxiang 453003, Henan, Peoples R China

[3] Jinan Univ, Shenzhen Eye Hosp, Shenzhen Eye Inst, 18 Zetian Rd, Shenzhen 518000, Peoples R China

[4] Ningxia Med Univ, Sch Basic Med Sci, Ningxia Key Lab Prevent & Control Common Infect Di, Yinchuan, Peoples R China

[5] Shanxi Med Univ, Hosp 3, Tongji Shanxi Hosp, Shanxi Acad Med Sci,Shanxi Bethune Hosp,Dept Ophth, N99 Longcheng St, Taiyuan 030032, Shanxi, Peoples R China

来源：

CRISPR JOURNAL | 2024年 / 7卷 / 04期

基金：

中国国家自然科学基金;

关键词：

GENE-THERAPY; EXPRESSION; EFFICIENT; VECTORS;

D O I：

10.1089/crispr.2024.0019

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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.

引用

页码：188 / 196

页数：9

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