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.
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.
Mol Ther. 2023 Jul 5;31(7):2286-2295. doi: 10.1016/j.ymthe.2023.02.015. Epub 2023 Feb 18.
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.
天使综合征（AS）是一种罕见的神经发育障碍，由母源表达的 UBE3A 功能丧失突变引起。到目前为止，患者还没有特定的基因治疗方法。尽管父源 UBE3A 是完整且转录活跃的，但在神经元中，通过反义长非编码 RNA UBE3A-ATS 的延伸使其沉默。在这里，我们证明了使用高保真 CRISPR-Cas13（hfCas13x.1）系统靶向父源 Ube3a-ATS 的 RNA 可以将 Ube3a 的表达恢复到与母源 Ube3a 相似的水平在培养的小鼠神经元中。此外，通过侧脑室注射用神经元特异性 hSyn1 启动子驱动的 hfCas13x.1 包装的腺相关病毒（AAV-PHP.eb）可以在治疗后长达 4 个月恢复新生 AS 小鼠皮质和海马体中的父源 Ube3a 表达。行为测试表明，父源 Ube3a 的表达显著缓解了 AS 相关症状，包括肥胖和运动功能。我们的研究结果表明，hfCas13x.1 介导的 Ube3a-ATS lncRNA 的抑制可能为 AS 提供了一种有前途的靶向干预措施。
