Treatment of autosomal dominant retinitis pigmentosa caused by RHO-P23H mutation with high-fidelity Cas13X in mice

Zixiang Yan; Yuqin Yao; Luyao Li; Lingqiong Cai; Haiwei Zhang; Shenghai Zhang; Qingquan Xiao; Xing Wang; Erwei Zuo; Chunlong Xu; Jihong Wu; Hui Yang

doi:10.1016/j.omtn.2023.08.002

Treatment of autosomal dominant retinitis pigmentosa caused by RHO-P23H mutation with high-fidelity Cas13X in mice

Mol Ther Nucleic Acids. 2023 Aug 7:33:750-761. doi: 10.1016/j.omtn.2023.08.002. eCollection 2023 Sep 12.

Authors

Zixiang Yan¹, Yuqin Yao², Luyao Li², Lingqiong Cai², Haiwei Zhang³, Shenghai Zhang^{4

5

6}, Qingquan Xiao², Xing Wang², Erwei Zuo¹, Chunlong Xu^{7

8}, Jihong Wu^{4

5

6}, Hui Yang^{2

3

7}

Affiliations

¹ Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Key Laboratory of Synthetic Biology, Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China.
² HuidaGene Therapeutics Co., Ltd., Shanghai 200131, China.
³ Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
⁴ Department of Ophthalmology, Eye and ENT Hospital, Shanghai Medical College, Fudan University, Shanghai 200032, China.
⁵ Shanghai Key Laboratory of Visual Impairment and Restoration, Science and Technology Commission of Shanghai Municipality, Shanghai, China.
⁶ Key Laboratory of Myopia (Fudan University), Chinese Academy of Medical Sciences, National Health Commission, Shanghai, China.
⁷ Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai 201210, China.
⁸ Lingang Laboratory, Shanghai 201210, China.

Abstract

Mutations in Rhodopsin (RHO) gene commonly cause autosomal dominant retinitis pigmentosa (adRP) without effective therapeutic treatment so far. Compared with genomic DNA-targeting CRISPR-Cas9 system, Cas13 edits RNA for therapeutic applications, avoiding the risk of causing permanent changes in the genome. In particular, a compact and high-fidelity Cas13X (hfCas13X) recently has been developed to degrade targeted RNA with minimal collateral effects and could also be packaged in a single adeno-associated virus for efficient in vivo delivery. In this study, we engineered single-guide RNA for hfCas13X to specifically knock down human mutant Rhodopsin transcripts RHO-P23H with minimal effect on wild-type transcripts. Moreover, treatment with hfCas13X alleviated the adRP progression in both RHO-P23H overexpression-induced and humanized hRHO^P23H/WT mouse models. Our study indicates the potential of hfCas13X in treating adRP caused by RHO mutations and other genetic diseases.

Keywords: MT: RNA/DNA editing; P23H mutation; adRP; hfCas13X; retinitis pigmentosa; rhodopsin.