Aksoy Yagiz Alp, Cole Alexander J, Deng Wei, Hesselson Daniel
Sydney Medical School, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2006, Australia.
Cancer Diagnosis and Pathology Group, Kolling Institute of Medical Research, Royal North Shore Hospital, St Leonards, NSW 2065, Australia.
Cells. 2024 Feb 21;13(5):372. doi: 10.3390/cells13050372.
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease characterized by the degeneration of motor neurons. Mutations in the cyclin F () and fused in sarcoma () genes have been associated with ALS pathology. In this study, we aimed to investigate the functional role of CCNF and FUS in ALS by using genome editing techniques to generate zebrafish models with genetic disruptions in these genes. Sequence comparisons showed significant homology between human and zebrafish CCNF and FUS proteins. We used CRISPR/Cas9 and TALEN-mediated genome editing to generate targeted disruptions in the zebrafish and genes. Ccnf-deficient zebrafish exhibited abnormal motor neuron development and axonal outgrowth, whereas Fus-deficient zebrafish did not exhibit developmental abnormalities or axonopathies in primary motor neurons. However, Fus-deficient zebrafish displayed motor impairments in response to oxidative and endoplasmic reticulum stress. The Ccnf-deficient zebrafish were only sensitized to endoplasmic reticulum stress, indicating that ALS genes have overlapping as well as unique cellular functions. These zebrafish models provide valuable platforms for studying the functional consequences of CCNF and FUS mutations in ALS pathogenesis. Furthermore, these zebrafish models expand the drug screening toolkit used to evaluate possible ALS treatments.
肌萎缩侧索硬化症（ALS）是一种毁灭性的神经退行性疾病，其特征是运动神经元退化。细胞周期蛋白F（CCNF）和肉瘤融合蛋白（FUS）基因的突变与ALS病理相关。在本研究中，我们旨在通过使用基因组编辑技术生成这些基因发生遗传破坏的斑马鱼模型，来研究CCNF和FUS在ALS中的功能作用。序列比较显示人类和斑马鱼的CCNF及FUS蛋白之间存在显著同源性。我们使用CRISPR/Cas9和TALEN介导的基因组编辑在斑马鱼的CCNF和FUS基因中产生靶向破坏。CCNF缺陷型斑马鱼表现出运动神经元发育异常和轴突生长异常，而FUS缺陷型斑马鱼在初级运动神经元中未表现出发育异常或轴突病变。然而，FUS缺陷型斑马鱼在氧化应激和内质网应激反应中表现出运动障碍。CCNF缺陷型斑马鱼仅对内质网应激敏感，这表明ALS相关基因具有重叠以及独特的细胞功能。这些斑马鱼模型为研究CCNF和FUS突变在ALS发病机制中的功能后果提供了有价值的平台。此外，这些斑马鱼模型扩展了用于评估可能的ALS治疗方法的药物筛选工具。
