Publication Date:
2019
Description:
〈p〉Leber congenital amaurosis (LCA), one of the leading causes of childhood-onset blindness, is caused by autosomal recessive mutations in several genes including 〈i〉RPE65〈/i〉. In this study, we performed CRISPR-Cas9–mediated therapeutic correction of a disease-associated nonsense mutation in 〈i〉Rpe65〈/i〉 in 〈i〉rd12〈/i〉 mice, a model of human LCA. Subretinal injection of adeno-associated virus carrying CRISPR-Cas9 and donor DNA resulted in 〉1% homology-directed repair and ~1.6% deletion of the pathogenic stop codon in 〈i〉Rpe65〈/i〉 in retinal pigment epithelial tissues of 〈i〉rd12〈/i〉 mice. The a- and b-waves of electroretinograms were recovered to levels up to 21.2 ± 4.1% and 39.8 ± 3.2% of their wild-type mice counterparts upon bright stimuli after dark adaptation 7 months after injection. There was no definite evidence of histologic perturbation or tumorigenesis during 7 months of observation. Collectively, we present the first therapeutic correction of an 〈i〉Rpe65〈/i〉 nonsense mutation using CRISPR-Cas9, providing new insight for developing therapeutics for LCA.〈/p〉
Electronic ISSN:
2375-2548
Topics:
Natural Sciences in General
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