CRISPR/Cas9 Delivery Mediated with Hydroxyl‐Rich Nanosystems for Gene Editing in Aorta. Issue 12 (20th April 2019)

• Record Type: Journal Article
• Authors: Zhang, Xiaoping; Xu, Chen; Gao, Shijuan; Li, Ping; Kong, Yu; Li, Tiantian; Li, Yulin; Xu, Fu‐Jian; Du, Jie
• Is Part Of: Advanced science. Volume 6:Issue 12(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-04-20
• Subjects: aorta disease -- cationic carriers -- CRISPR‐associated nuclease 9 delivery -- genome editing -- hydroxyl‐rich; Science -- Periodicals; 505
• DOI: 10.1002/advs.201900386
• Languages: English
• ISSNs: 2198-3844

Abstract

Abstract: A CRISPR/Cas9 system has emerged as a powerful tool for gene editing to treat genetic mutation related diseases. Due to the complete endothelial barrier, effective delivery of the CRISPR/Cas9 system to vasculatures remains a challenge for in vivo gene editing of genetic vascular diseases especially in aorta. Herein, it is reported that CHO‐PGEA (cholesterol (CHO)‐terminated ethanolamine‐aminated poly(glycidyl methacrylate)) with rich hydroxyl groups can deliver a plasmid based pCas9‐sg Fbn 1 system for the knockout of exon 10 in Fbn 1 gene. This is the first report of a polycation‐mediated CRISPR/Cas9 system for gene editing in aorta of adult mice. CHO‐PGEA/pCas9‐sg Fbn 1 nanosystems can effectively contribute to the knockout of exon 10 in Fbn 1 in vascular smooth muscle cells in vitro, which leads to the change of the phosphorylation of Smad2/3 and the increased expression of two downstream signals of Fbn 1: Mmp‐ 2 and Ctgf . For in vivo application, the aortic enrichment of CHO‐PGEA/Cas9‐sg Fbn 1 is achieved by administering a pressor dose of angiotensin II (Ang II). The effects of the pCas9‐sg Fbn 1 system targeting Fbn 1 demonstrate an increase in the expression of Mmp‐ 2 and Ctgf in aorta. Thus, the combination of CHO‐PGEA/pCas9‐sg Fbn 1 nanosystems with Ang II infusion can provide the possibility for in vivo gene editing in aorta. Abstract : In this work, cholesterol (CHO)‐terminated ethanolamine‐aminated poly(glycidyl methacrylate) (CHO‐PGEA) with rich hydroxyl groups is applied to efficiently deliver CRISPR/Cas9 plasmid for gene editing in adult mouse aorta. Ang II infusion significantly increases the enrichment of CHO‐PGEA/pCas9‐sg Fbn 1 in aortic tissues. CHO‐PGEA/pCas9‐sg Fbn 1 can significantly induce loss‐of‐function mutations of the Fbn 1 gene and cause aortic diameter dilation.