Creation of de novo cryptic splicing for ALS/FTD precision medicine.
Creation of de novo cryptic splicing for ALS/FTD precision medicine.
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为 ALS/FTD 精准医学创建从头神秘剪接。
DOI：
10.1101/2023.11.15.565967
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Kard
中科院分区：
文献类型：
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作者：
Wilkins,OscarG;Chien,MaxZYJ;Wlaschin,JosetteJ;Pisliakova,Maria;Thompson,David;Digby,Holly;Simkin,RebeccaL;Diaz,JuanAntinao;Mehta,PujaR;Keuss,MatthewJ;Zanovello,Matteo;Brown,Anna-Leigh;Harley,Peter;Darbey,Annalucia;Kard
RNA结合蛋白TDP-43(TDP-LOF)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诱导的隐蔽剪接的特异性来驱动蛋白质在疾病过程发生的时间和地点的表达。SpliceNouveau算法将深度学习与理性设计相结合，在蛋白质编码序列中生成可定制的隐蔽剪接事件。我们证明了TDP-Reg报告基因在体外和体内的表达与TDP-LOF紧密相连。TDP-Reg能够对基因组进行编辑，以特异性地在TDP-LOF上去除UNC13A隐秘的供体剪接位点。最后，我们设计了编码TDP-43/Rver1融合蛋白的TDP-Reg载体，以挽救关键的病理隐蔽剪接事件，为TDP43相关疾病的精确治疗的发展铺平了道路。
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.