AAV-mediated In vivo genome editing in vascular endothelial cells
AAV-mediated In vivo genome editing in vascular endothelial cells
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AAV 介导的血管内皮细胞体内基因组编辑
DOI：
10.1016/j.ymeth.2020.12.001
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发表时间：
2020
期刊：
影响因子：
4.8
通讯作者：
Lei Hetian
中科院分区：
文献类型：
--
作者：
Wu Wenyi;Yang Yanhui;Yao Fei;Dong Lijun;Xia Xiaobo;Zhang Shaochong;Lei Hetian
体内基因组编辑面临许多挑战，包括效率和安全性。在这里，我们描述了一种用腺相关病毒（AAV）将CRISPR-Cas9递送到血管内皮细胞中的有效体内基因组编辑方法。在该系统中，SpCas 9的表达由细胞间粘附分子2（pICAM 2）的特异性内皮启动子驱动，以将这种外源酶限制在血管内皮细胞中，其可以被AAV 1有效感染。我们通过在氧诱导的视网膜病变小鼠模型中编辑视网膜血管内皮细胞中的VEGFR 2来验证这种方法，并期望这种简化的方案可以扩展到体内编辑内皮基因组的其他研究。
In vivogenome editing meets numerous challenges including efficiency and safety. Here we describe an efficient in vivo genome editing method of delivering CRISPR-Cas9 into vascular endothelial cells with adeno-associated viruses (AAVs). In this system, expression of SpCas9 is driven by a specific endothelial promoter of intercellular adhesion molecule 2 (pICAM2) to restrict this foreign enzyme in vascular endothelial cells, which can be efficiently infected by AAV1. We exemplify this approach by editingVEGFR2in retinal vascular endothelial cells in a mouse model of oxygen-induced retinopathy, and expect that this simplified protocol can be expanded to other researches on editing endothelial genome in vivo.