CRISPR/Cas9 Delivery Mediated with Hydroxyl‐Rich Nanosystems for Gene Editing in Aorta

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.

Nanoparticles‐Mediated CRISPR/Cas Gene Editing Delivery System

“…Zhang et al [63] constructed a dual-locking nanoparticle (DLNP) that can restrict CRISPR/Cas13a activation to tumor tissues. This polymer layer endows the DLNP with enhanced stability during blood circulation or in normal tissues, and facilitates cellular internalization of the CRISPR/Cas13a system and activation of gene editing upon entry into tumor tissue.…”

The Important Role of Endothelium and Extracellular Vesicles in the Cellular Mechanism of Aortic Aneurysm Formation

“…In addition to standard pharmacological and surgical therapy, biomarkers and stem cells are also widely used. It is noteworthy that it is possible to apply genome editing technology based on CRISPR/Cas9 in the targeted therapy, which seems to be a promising future for treating aortic aneurysms [ 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 ].…”

A Lactose‐Derived CRISPR/Cas9 Delivery System for Efficient Genome Editing In Vivo to Treat Orthotopic Hepatocellular Carcinoma

“…However, the clinical translation potential of viral vectors remains limited, mostly due to the possible long-term toxicity associated with the risk of integrating virus sequence into human genome. [11] Non-virus delivery of the CRISPR/Cas9 system might be a promising way for its therapeutic application, [12][13][14][15][16][17] especially considering the difficulty of packing the Cas9 gene with large size (about 4.2 kb) into a viral vector. Additionally, the employment of non-virus vectors would significantly decline the risk of integrating virus sequence into human genome.…”