CRISPR/Cas9-Mediated Excision of ALS/FTD-Causing Hexanucleotide Repeat Expansion in C9ORF72 rescues major disease mechanisms in vivo and in vitro
CRISPR/Cas9-Mediated Excision of ALS/FTD-Causing Hexanucleotide Repeat Expansion in C9ORF72 rescues major disease mechanisms in vivo and in vitro
Abstract A hexanucleotide repeat expansion (HRE) consisting of GGGGCC24+ in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Both are fatal neurodegenerative diseases with no current approved treatments that significantly slow disease progression or extend life expectancy. Several hypotheses have emerged to explain how this HRE causes neuronal death, including C9ORF72 haploinsufficiency, sequestration of RNA-binding proteins in the nucleus, and production of dipeptide repeat proteins. In the present study we used a CRISPR/Cas9 gene-editing approach to remove the HRE from the C9ORF72 genomic locus, designing guide RNAs (gRNAs) flanking the HRE, and delivered Cas9 and gRNAs using adeno-associated virus serotype 9 (AAV9) vectors. Here, we demonstrate successful excision of the HRE in C9ORF72 in primary cortical neurons and brains of three mouse models containing the C9ORF72 expanded HRE (ranging from 500-600 repeats) as well as in patient-derived iPSC motor neurons and brain organoids (450 repeats). This resulted in a reduction of RNA foci, poly-dipeptides and haploinsufficiency, the major hallmarks of C9-ALS/FTD, making this an extremely attractive therapeutic approach to these diseases.
Edraki Alireza、Kraus Carolyn、Krishnan Gopinath、Esanov Rustam、Andrade Nadja S.、Rybin Matthew J.、Ramic Melina、Stephens Zachary D.、Kahriman Aydan、Benatar Michael、Brodsky Michael H.、Petrucelli Leonard、Gao Fen-Biao、Sontheimer Erik J.、Brown Robert H.、Abdallah Abbas、Mueller Christian、Rodriguez Tom¨￠s、Nagase Hiroko、Kocher Jean-Pierre A.、Meijboom Katharina E.、Fordham Nicholas P.、Gendron Tania F.、Henninger Nils、Blackwood Meghan T.、Zeier Zane
RNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical SchoolRNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical SchoolDepartment of Neurology, University of Massachusetts Medical SchoolDepartment of Psychiatry & Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of MedicineDepartment of Psychiatry & Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of MedicineDepartment of Psychiatry & Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of MedicineDepartment of Psychiatry & Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of MedicineDepartment of Quantitative Health Sciences. Mayo ClinicDepartment of Neurology, University of Massachusetts Medical SchoolDepartment of Neurology, University of Miami Miller School of MedicineDepartment of Molecular, Cell and Cancer Biology, University of Massachusetts Medical SchoolDepartment of Neuroscience, Mayo ClinicDepartment of Neurology, University of Massachusetts Medical SchoolRNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical SchoolDepartment of Neurology, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolRNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolDepartment of Quantitative Health Sciences. Mayo ClinicHorae Gene Therapy Center, University of Massachusetts Medical School||Department of Neurology, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolDepartment of Neuroscience, Mayo ClinicDepartment of Neurology, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolDepartment of Psychiatry & Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of Medicine||RNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical School
神经病学、精神病学基础医学生物科学研究方法、生物科学研究技术
Edraki Alireza,Kraus Carolyn,Krishnan Gopinath,Esanov Rustam,Andrade Nadja S.,Rybin Matthew J.,Ramic Melina,Stephens Zachary D.,Kahriman Aydan,Benatar Michael,Brodsky Michael H.,Petrucelli Leonard,Gao Fen-Biao,Sontheimer Erik J.,Brown Robert H.,Abdallah Abbas,Mueller Christian,Rodriguez Tom¨￠s,Nagase Hiroko,Kocher Jean-Pierre A.,Meijboom Katharina E.,Fordham Nicholas P.,Gendron Tania F.,Henninger Nils,Blackwood Meghan T.,Zeier Zane.CRISPR/Cas9-Mediated Excision of ALS/FTD-Causing Hexanucleotide Repeat Expansion in C9ORF72 rescues major disease mechanisms in vivo and in vitro[EB/OL].(2025-03-28)[2025-08-02].https://www.biorxiv.org/content/10.1101/2022.05.17.492303.点此复制
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