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AAV-mediated in vivo genome editing in vascular endothelial cells.

Authors
  • Wu, Wenyi1
  • Yang, Yanhui2
  • Yao, Fei1
  • Dong, Lijun3
  • Xia, Xiaobo4
  • Zhang, Shaochong5
  • Lei, Hetian6
  • 1 Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, PR China; Hunan Key Laboratory of Ophthalmology, Changsha, PR China. , (China)
  • 2 School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, Ningxia, PR China. , (China)
  • 3 Shenzhen Eye Institute, Shenzhen Eye Hospital, Jinan University, Shenzhen, PR China. , (China)
  • 4 Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, PR China; Hunan Key Laboratory of Ophthalmology, Changsha, PR China. Electronic address: [email protected] , (China)
  • 5 Shenzhen Eye Institute, Shenzhen Eye Hospital, Jinan University, Shenzhen, PR China. Electronic address: [email protected] , (China)
  • 6 Shenzhen Eye Institute, Shenzhen Eye Hospital, Jinan University, Shenzhen, PR China. Electronic address: [email protected] , (China)
Type
Published Article
Journal
Methods
Publisher
Elsevier
Publication Date
Oct 01, 2021
Volume
194
Pages
12–17
Identifiers
DOI: 10.1016/j.ymeth.2020.12.001
PMID: 33309782
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

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. Copyright © 2020. Published by Elsevier Inc.

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