Creation of de novo cryptic splicing for ALS and FTD precision medicine

• 成果类型: Article
• 署名作者: Wilkins, Oscar G.; Chien, Max Z. Y. J.; Wlaschin, Josette J.; Barattucci, Simone; Harley, Peter; Mattedi, Francesca; Mehta, Puja R.; Pisliakova, Maria; Ryadnov, Eugeni; Keuss, Matthew J.; Thompson, David; Digby, Holly; Knez, Lea; Simkin, Rebecca L.; Diaz, Juan Antinao; Zanovello, Matteo; Brown, Anna-Leigh; Darbey, Annalucia; Karda, Rajvinder; Fisher, Elizabeth M. C.; Cunningham, Thomas J.; Le Pichon, Claire E.; Ule, Jernej; Fratta, Pietro
• 署名单位: University of London; University College London; Francis Crick Institute; National Institutes of Health (NIH) - USA; NIH Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD); Johns Hopkins University; MRC Harwell; University of London; King's College London; University of London; University College London; University of London; University College London; University of London; University College London
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-10951
• DOI: 10.1126/science.adk2539
• 发表日期: 2024-10-04
• 页码: 61-69
• 关键词: tdp-43

摘要

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.