Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
Nat Commun. 2023 Oct 14;14(1):6492. doi: 10.1038/s41467-023-42147-z.
The TDP-43 proteinopathies, which include amyotrophic lateral sclerosis and frontotemporal dementia, are a devastating group of neurodegenerative disorders that are characterized by the mislocalization and aggregation of TDP-43. Here we demonstrate that RNA-targeting CRISPR effector proteins, a programmable class of gene silencing agents that includes the Cas13 family of enzymes and Cas7-11, can be used to mitigate TDP-43 pathology when programmed to target ataxin-2, a modifier of TDP-43-associated toxicity. In addition to inhibiting the aggregation and transit of TDP-43 to stress granules, we find that the in vivo delivery of an ataxin-2-targeting Cas13 system to a mouse model of TDP-43 proteinopathy improved functional deficits, extended survival, and reduced the severity of neuropathological hallmarks. Further, we benchmark RNA-targeting CRISPR platforms against ataxin-2 and find that high-fidelity forms of Cas13 possess improved transcriptome-wide specificity compared to Cas7-11 and a first-generation effector. Our results demonstrate the potential of CRISPR technology for TDP-43 proteinopathies.
TDP-43 蛋白病包括肌萎缩性侧索硬化症和额颞叶痴呆，是一组具有破坏性的神经退行性疾病，其特征是 TDP-43 的定位和聚集错误。在这里，我们证明 RNA 靶向 CRISPR 效应蛋白，包括 Cas13 酶家族和 Cas7-11 的可编程基因沉默剂类，可以被用来减轻 TDP-43 病理，当被编程靶向 ataxin-2 时，ataxin-2 是 TDP-43 相关毒性的修饰因子。除了抑制 TDP-43 聚集和向应激颗粒的转运外，我们还发现，将靶向 ataxin-2 的 Cas13 系统递送到 TDP-43 蛋白病的小鼠模型中，可以改善功能缺陷，延长存活时间，并减轻神经病理学特征的严重程度。此外，我们将 RNA 靶向 CRISPR 平台与 ataxin-2 进行基准测试，发现与 Cas7-11 和第一代效应物相比，高保真形式的 Cas13 具有改善的全转录组特异性。我们的结果表明 CRISPR 技术在 TDP-43 蛋白病中的应用潜力。
