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.

Angelman 综合征 (AS) 是一种罕见的神经发育障碍，由母体表达的UBE3A功能缺失突变引起。迄今为止，还没有针对患者的基因特异性治疗。尽管完整且具有转录活性，但父系遗传的UBE3A通过神经元中反义长非编码 RNA UBE3A-ATS的延伸而被沉默。在这里，我们证明，用高保真 CRISPR-Cas13 (hfCas13x.1) 系统对父本Ube3a-ATS进行 RNA 靶向，可以将培养的小鼠神经元中的Ube3a表达恢复到与母本Ube3a相似的水平。此外，将腺相关病毒（AAV-PHP.eb）中包装的神经元特异性hSyn1启动子驱动的 hfCas13x.1 注射到侧脑室，可以在治疗后长达 4 个月的时间内恢复新生 AS 小鼠皮层和海马中父本Ube3a的表达。行为测试表明，父亲Ube3a的表达显着减轻了 AS 相关症状，包括肥胖和运动功能。我们的结果表明，hfCas13x.1 介导的Ube3a-ATS lncRNA 抑制可能成为 AS 的一种有前途的靶向干预措施。