UCL Queen Square Motor Neuron Disease Centre, Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology, University College London, London WC1N 3BG, UK.
The Francis Crick Institute, London NW1 1AT, UK.
Science. 2024 Oct 4;386(6717):61-69. doi: 10.1126/science.adk2539. Epub 2024 Oct 3.
Loss of function of the RNA-binding protein TDP-43 (TDP-LOF) is a hallmark of amyotrophic lateral sclerosis (ALS) and other neurodegenerative disorders. Here we describe TDP-REG, which exploits the specificity of cryptic splicing induced by TDP-LOF to drive protein expression when and where the disease process occurs. The SpliceNouveau algorithm combines deep learning with rational design to generate customizable cryptic splicing events within protein-coding sequences. We demonstrate that expression of TDP-REG reporters is tightly coupled to TDP-LOF in vitro and in vivo. TDP-REG enables genomic prime editing to ablate the UNC13A cryptic donor splice site specifically upon TDP-LOF. Finally, we design TDP-REG vectors encoding a TDP-43/Raver1 fusion protein that rescues key pathological cryptic splicing events, paving the way for the development of precision therapies for TDP43-related disorders.
TDP-43（TDP-LOF）的 RNA 结合蛋白功能丧失是肌萎缩侧索硬化症（ALS）和其他神经退行性疾病的标志。在这里，我们描述了 TDP-REG，它利用 TDP-LOF 诱导的隐秘剪接的特异性，在疾病发生的时间和地点驱动蛋白质表达。SpliceNouveau 算法将深度学习与合理设计相结合，在蛋白质编码序列中产生可定制的隐秘剪接事件。我们证明，TDP-REG 报告基因的表达与体外和体内的 TDP-LOF 紧密相关。TDP-REG 使基因组原位编辑能够在 TDP-LOF 时特异性地切除 UNC13A 隐秘供体位点。最后，我们设计了 TDP-REG 载体，编码 TDP-43/Raver1 融合蛋白，可挽救关键的病理性隐秘剪接事件，为 TDP43 相关疾病的精准治疗铺平道路。
