Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy

eLife

Jayakrishna Ambati¹  Felipe Pereira¹  Siddharth Narendran¹  Wei Liu²  Yue Zhang²  Guangping Gao³  Sai Bhuvanagiri⁴  Susie Choi⁴  Austin Bohner⁴  Jinlu Liu⁴  Lara Carroll⁴  Balamurali K Ambati⁵  Hironori Uehara⁵  Sangeetha Ravi Kumar⁵  Bonnie Archer⁵  Xiaohui Zhang⁵  Albert S Jun⁶ 

[1] Department of Ophthalmology, University of Virginia, Charlottesville, United States;Division of Epidemiology, Department of Internal Medicine, University of Utah, Salt Lake City, United States;Gene Therapy Center, Department of Microbiology and Physiological Science Systems, University of Massachusetts Medical School, Worcester, United States;Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States;Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States;Wilmer Eye Institute, Johns Hopkins University, Baltimore, United States;

关键词: Fuchs endothelial corneal dystrophy;    CRISPR/Cas9;    Adenovirus;    off-target analysis of CRISPR/Cas9;    Human;    Mouse;   

DOI  :  10.7554/eLife.55637

来源: eLife Sciences Publications, Ltd

【 摘 要 】

A missense mutation of collagen type VIII alpha 2 chain (COL8A2) gene leads to early-onset Fuchs’ endothelial corneal dystrophy (FECD), which progressively impairs vision through the loss of corneal endothelial cells. We demonstrate that CRISPR/Cas9-based postnatal gene editing achieves structural and functional rescue in a mouse model of FECD. A single intraocular injection of an adenovirus encoding both the Cas9 gene and guide RNA (Ad-Cas9-Col8a2gRNA) efficiently knocked down mutant COL8A2 expression in corneal endothelial cells, prevented endothelial cell loss, and rescued corneal endothelium pumping function in adult Col8a2 mutant mice. There were no adverse sequelae on histology or electroretinography. Col8a2 start codon disruption represents a non-surgical strategy to prevent vision loss in early-onset FECD. As this demonstrates the ability of Ad-Cas9-gRNA to restore the phenotype in adult post-mitotic cells, this method may be widely applicable to adult-onset diseases, even in tissues affected with disorders of non-reproducing cells.

【 授权许可】

CC BY