Journal
METHODS
Volume 194, Issue -, Pages 12-17Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2020.12.001
Keywords
CRISPR-Cas9; AAV1; Vascular endothelial cells; A mouse model of oxygen-induced retinopathy
Funding
- Science and Technology Plan Project of Hunan Province [2019RS2011]
- National Natural Science Foundation of China [81900893, 81974134, 82070989]
- Sanming Project of Medicine in Shenzhen [SZSM202011015]
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This method delivers CRISPR-Cas9 into vascular endothelial cells using AAVs, with SpCas9 expression driven by a specific endothelial promoter to restrict the foreign enzyme in these cells. By editing VEGFR2 in retinal vascular endothelial cells in a mouse model, this simplified protocol can potentially be expanded to other research on editing endothelial genome in vivo.
In vivo genome 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 editing VEGFR2 in 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.
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