A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice

Jinhui Li; Zhixin Shen; Yajing Liu; Zixiang Yan; Yuanhua Liu; Xiang Lin; Junjie Tang; Ruimin Lv; Guannan Geng; Zhi-Qi Xiong; Changyang Zhou; Hui Yang

doi:10.1016/j.ymthe.2023.02.015

A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice

Mol Ther. 2023 Jul 5;31(7):2286-2295. doi: 10.1016/j.ymthe.2023.02.015. Epub 2023 Feb 18.

Authors

Jinhui Li¹, Zhixin Shen¹, Yajing Liu¹, Zixiang Yan², Yuanhua Liu¹, Xiang Lin³, Junjie Tang¹, Ruimin Lv¹, Guannan Geng¹, Zhi-Qi Xiong⁴, Changyang Zhou⁵, Hui Yang⁶

Affiliations

¹ 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.
² Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China.
³ Department of Neurology and Institute of Neurology, The First Affiliated Hospital, Institute of Neuroscience, Fujian Medical University, #20, Chazhong Road, Taijiang District, Fuzhou 350005, China.
⁴ 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. Electronic address: xiongzhiqi@ion.ac.cn.
⁵ 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. Electronic address: zhouchangyang@ion.ac.cn.
⁶ 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; Shanghai Center for Brain Science and Brain-Inspired Intelligence Technology, Shanghai 201210, China; HuidaGene Therapeutics Co., Ltd., Shanghai 200131, China. Electronic address: huiyang@ion.ac.cn.

PMID: 36805082
PMCID: PMC10362381 (available on 2024-07-05)
DOI: 10.1016/j.ymthe.2023.02.015

Abstract

Angelman syndrome (AS) is a rare neurodevelopmental disorder caused by loss of function mutations in maternally expressed UBE3A. No gene-specific treatment is available for patients so far. Although intact and transcriptionally active, paternally inherited UBE3A is silenced by elongation of antisense long noncoding RNA UBE3A-ATS in neurons. Here, we demonstrated that RNA targeting of paternal Ube3a-ATS with a high-fidelity CRISPR-Cas13 (hfCas13x.1) system could restore Ube3a expression to similar levels as that of maternal Ube3a in the cultured mouse neurons. Furthermore, injection into lateral ventricles with neuron-specific hSyn1 promoter-driven hfCas13x.1 packaged in adeno-associated virus (AAV-PHP.eb) could restore paternal Ube3a expression in cortex and hippocampus of neonatal AS mice for up to 4 months after treatment. Behavioral tests showed that expression of paternal Ube3a significantly alleviated AS-related symptoms, including obesity and motor function. Our results suggested that hfCas13x.1-mediated suppression of the Ube3a-ATS lncRNA potentially serves as a promising targeted intervention for AS.

Keywords: Angelman syndrome; RNA editing; high-fidelity Cas13; motor function; paternal Ube3a.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angelman Syndrome* / genetics
Angelman Syndrome* / therapy
Animals
Mice
Obesity
RNA, Antisense / genetics
Ubiquitin-Protein Ligases / genetics

Substances

RNA, Antisense
Ubiquitin-Protein Ligases
Ube3a protein, mouse