CRISPR/Cas9-mediated excision of ALS/FTD-causing hexanucleotide repeat expansion in C9ORF72 rescues major disease mechanisms in vivo and in vitro
Meijboom, Katharina E ; Abdallah, Abbas ; Fordham, Nicholas P ; Nagase, Hiroko ; Rodriguez, Tomás ; Kraus, Carolyn ; Gendron, Tania F ; Krishnan, Gopinath ; Esanov, Rustam ; Andrade, Nadja S ... show 10 more
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Abdallah, Abbas
Fordham, Nicholas P
Nagase, Hiroko
Rodriguez, Tomás
Kraus, Carolyn
Gendron, Tania F
Krishnan, Gopinath
Esanov, Rustam
Andrade, Nadja S
Rybin, Matthew J
Ramic, Melina
Stephens, Zachary D
Edraki, Alireza
Blackwood, Meghan
Kahriman, Aydan
Henninger, Nils
Kocher, Jean-Pierre A
Benatar, Michael
Brodsky, Michael H
Petrucelli, Leonard
Gao, Fen-Biao
Sontheimer, Erik J
Brown, Robert H
Zeier, Zane
Mueller, Christian
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Abstract
A GGGGCC24+ hexanucleotide repeat expansion (HRE) in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), fatal neurodegenerative diseases with no cure or approved treatments that substantially slow disease progression or extend survival. Mechanistic underpinnings of neuronal death include C9ORF72 haploinsufficiency, sequestration of RNA-binding proteins in the nucleus, and production of dipeptide repeat proteins. Here, we used an adeno-associated viral vector system to deliver CRISPR/Cas9 gene-editing machineries to effectuate the removal of the HRE from the C9ORF72 genomic locus. We demonstrate successful excision of the HRE in primary cortical neurons and brains of three mouse models containing the expansion (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, major hallmarks of C9-ALS/FTD, making this a promising therapeutic approach to these diseases.
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Meijboom KE, Abdallah A, Fordham NP, Nagase H, Rodriguez T, Kraus C, Gendron TF, Krishnan G, Esanov R, Andrade NS, Rybin MJ, Ramic M, Stephens ZD, Edraki A, Blackwood MT, Kahriman A, Henninger N, Kocher JA, Benatar M, Brodsky MH, Petrucelli L, Gao FB, Sontheimer EJ, Brown RH, Zeier Z, Mueller C. CRISPR/Cas9-mediated excision of ALS/FTD-causing hexanucleotide repeat expansion in C9ORF72 rescues major disease mechanisms in vivo and in vitro. Nat Commun. 2022 Oct 21;13(1):6286. doi: 10.1038/s41467-022-33332-7. PMID: 36271076; PMCID: PMC9587249.