Epilepsy is a major health burden, calling for new mechanistic insights and therapies. CRISPR-mediated gene editing shows promise to cure genetic pathologies, although hitherto it has mostly been applied ex vivo. Its translational potential for treating non-genetic pathologies is still unexplored. Furthermore, neurological diseases represent an important challenge for the application of CRISPR, because of the need in many cases to manipulate gene function of neurons in situ. A variant of CRISPR, CRISPRa, offers the possibility to modulate the expression of endogenous genes by directly targeting their promoters. We asked if this strategy can effectively treat acquired focal epilepsy, focusing on ion channels because their manipulation is known be effective in changing network hyperactivity and hypersynchronziation. We applied a doxycycline-inducible CRISPRa technology to increase the expression of the potassium channel gene Kcna1 (encoding Kv1.1) in mouse hippocampal excitatory neurons. CRISPRa-mediated Kv1.1 upregulation led to a substantial decrease in neuronal excitability. Continuous video-EEG telemetry showed that AAV9-mediated delivery of CRISPRa, upon doxycycline administration, decreased spontaneous generalized tonic-clonic seizures in a model of temporal lobe epilepsy, and rescued cognitive impairment and transcriptomic alterations associated with chronic epilepsy. The focal treatment minimizes concerns about off-target effects in other organs and brain areas. This study provides the proof-of-principle for a translational CRISPR-based approach to treat neurological diseases characterized by abnormal circuit excitability.

中文翻译：

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体内 CRISPRa 在啮齿动物癫痫模型中减少癫痫发作并挽救认知缺陷。

癫痫是一个主要的健康负担，需要新的机制见解和疗法。CRISPR 介导的基因编辑有望治愈遗传疾病，尽管迄今为止它主要是在体外应用。其治疗非遗传病理学的转化潜力仍未开发。此外，神经系统疾病代表了 CRISPR 应用的一个重要挑战，因为在许多情况下需要原位操纵神经元的基因功能。CRISPR 的一种变体 CRISPRa 提供了通过直接靶向其启动子来调节内源基因表达的可能性。我们询问该策略是否可以有效治疗获得性局灶性癫痫，重点关注离子通道，因为已知它们的操作可有效改变网络过度活跃和超同步。我们应用强力霉素诱导的 CRISPRa 技术来增加小鼠海马兴奋性神经元中钾通道基因 Kcna1（编码 Kv1.1）的表达。CRISPRa 介导的 Kv1.1 上调导致神经元兴奋性显着降低。连续视频脑电图遥测显示，在给予强力霉素后，AAV9 介导的 CRISPRa 递送减少了颞叶癫痫模型中自发的全身强直-阵挛性癫痫发作，并挽救了与慢性癫痫相关的认知障碍和转录组学改变。焦点治疗最大限度地减少了对其他器官和大脑区域的脱靶效应的担忧。本研究为基于 CRISPR 的转化方法治疗以异常电路兴奋性为特征的神经系统疾病提供了原理验证。