TY - JOUR TI - Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy AU - Uehara, Hironori AU - Zhang, Xiaohui AU - Pereira, Felipe AU - Narendran, Siddharth AU - Choi, Susie AU - Bhuvanagiri, Sai AU - Liu, Jinlu AU - Ravi Kumar, Sangeetha AU - Bohner, Austin AU - Carroll, Lara AU - Archer, Bonnie AU - Zhang, Yue AU - Liu, Wei AU - Gao, Guangping AU - Ambati, Jayakrishna AU - Jun, Albert S AU - Ambati, Balamurali K A2 - Zoghbi, Huda Y A2 - Cepko, Constance L A2 - Ksander, Bruce VL - 10 PY - 2021 DA - 2021/06/08 SP - e55637 C1 - eLife 2021;10:e55637 DO - 10.7554/eLife.55637 UR - https://doi.org/10.7554/eLife.55637 AB - 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. KW - Fuchs endothelial corneal dystrophy KW - CRISPR/Cas9 KW - Adenovirus KW - off-target analysis of CRISPR/Cas9 JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -