Direct reprogramming based on genetic factors resembles a promising strategy to replace lost cells in degenerative diseases such as Parkinson's disease. For this, we developed a knock-in mouse line carrying a dual dCas9 transactivator system (dCAM) allowing the conditional in vivo activation of endogenous genes. To enable a translational application, we additionally established an AAV-based strategy carrying intein-split-dCas9 in combination with activators (AAV-dCAS). Both approaches were successful in reprogramming striatal astrocytes into induced GABAergic neurons confirmed by single-cell transcriptome analysis of reprogrammed neurons in vivo. These GABAergic neurons functionally integrate into striatal circuits, alleviating voluntary motor behavior aspects in a 6-OHDA Parkinson's disease model. Our results suggest a novel intervention strategy beyond the restoration of dopamine levels. Thus, the AAV-dCAS approach might enable an alternative route for clinical therapies of Parkinson's disease.

中文翻译：

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通过 CRISPRa 将星形胶质细胞重编程为 GABA 能神经元来拯救帕金森病运动症状。

基于遗传因素的直接重编程类似于替代帕金森病等退行性疾病中丢失的细胞的有希望的策略。为此，我们开发了一种带有双 dCas9 反式激活系统 (dCAM) 的敲入小鼠系，允许有条件地在体内激活内源基因。为了实现翻译应用，我们还建立了一种基于 AAV 的策略，该策略携带内含肽-分裂-dCas9 与激活剂 (AAV-dCAS) 结合。两种方法都成功地将纹状体星形胶质细胞重编程为诱导的 GABA 能神经元，这通过体内重编程神经元的单细胞转录组分析得到证实。这些 GABA 能神经元在功能上整合到纹状体回路中，减轻 6-OHDA 帕金森病模型中的自主运动行为方面。我们的研究结果表明，除了恢复多巴胺水平之外，还有一种新的干预策略。因此，AAV-dCAS 方法可能为帕金森病的临床治疗提供替代途径。