Disruption of splicing-regulatory elements using CRISPR/Cas9 to rescue spinal muscular atrophy in human iPSCs and mice
Disruption of splicing-regulatory elements using CRISPR/Cas9 to rescue spinal muscular atrophy in human iPSCs and mice作者机构：Department of Neurology and Institute of Neurology The First Affiliated Hospital of Fujian Medical University Fujian Key Laboratory of Molecular Neurology Fujian Medical University Institute of Neuroscience State Key Laboratory of Neuroscience Key Laboratory of Primate Neurobiology CAS Center for Excellence in Brain Science and Intelligence Technology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Key Lab of Computational Biology CAS-MPG Partner Institute for Computational Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Department of Anatomy Histology & Embryology Shanghai Medical College Fudan University Cold Spring Harbor LaboratoryCold Spring Harbor
出 版 物：《National Science Review》 (国家科学评论(英文版))
年 卷 期：2020年第7卷第1期
页 面：92-101页
核心收录：
学科分类：1002[医学-临床医学] 100204[医学-神经病学] 10[医学]
基 金：supported by the grant 81322017(W.-J.C.),81771230(W.-J.C.),31522037(H.Y.)and U1505222(N.W.)from the National Natural Science Foundation of China Joint Funds for the Innovation of Science and Technology of Fujian Province(2017Y9094)(W.-J.C.) National Key Clinical Specialty Discipline Construction Program(N.W.) Key Clinical Specialty Discipline Construction Program of Fujian(N.W.) National Science and Technology major project(2017YFC1001302)(H.Y.) Shanghai City Committee of Science and Technology project(16JC1420202)(H.Y.)
主 题：spinal muscular atrophy SMN2 splicing-regulatory elements CRISPR/Cas9 germline correction
摘 要：We here report a genome-editing strategy to correct spinal muscular atrophy(SMA). Rather than directly targeting the pathogenic exonic mutations, our strategy employed Cas9 and guide-sg RNA for the targeted disruption of intronic splicing-regulatory elements. We disrupted intronic splicing silencers(ISSs, including ISS-N1 and ISS + 100) of survival motor neuron(SMN) 2, a key modifier gene of SMA, to enhance exon 7 inclusion and full-length SMN expression in SMA iPSCs. Survival of splicing-corrected iPSC-derived motor neurons was rescued with SMN restoration. Furthermore, co-injection of Cas9 mRNA from Streptococcus pyogenes(SpCas9) or Cas9 from Staphylococcus aureus(SaCas9) alongside their corresponding sgRNAs targeting ISS-N1 into zygotes rescued 56% and 100% of severe SMA transgenic mice(Smn-/-, SMN2tg/-).The median survival of the resulting mice was extended to 400 days. Collectively, our study provides proof-of-principle for a new strategy to therapeutically intervene in SMA and other RNA-splicing-related diseases.
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