CRISPR deletion of the C9ORF72 promoter in ALS/FTD patient motor neurons abolishes production of dipeptide repeat proteins and rescues neurodegeneration
Gopinath Krishnan, Yu Zhang, Yuanzheng Gu, Mark W. Kankel, Fen‐Biao Gao, Sandra Almeida
Abstract
GGGGCC (G4C2) repeat expansion in the first intron of C9ORF72 is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Brain tissues from affected individuals show characteristic nuclear RNA foci containing the expanded repeat RNAs, as well as neuronal inclusions containing dipeptide repeat (DPR) proteins [poly(GA), poly(GP), poly(GR), poly(PR), and poly(PA)] resulting from the translation of both sense and antisense repeat RNAs in all reading frames. Although reduced C9ORF72 protein function may contribute to disease, the more likely drivers of disease are mechanisms related to a gain of toxic function. Currently, intense efforts are being made to identify disease mechanisms amenable for the development of therapeutic strategies. One promising avenue would be to prevent the production of the expanded repeat RNAs, such as by antisense oligonucleotides. Here, we tested another potential therapeutic approach: CRISPR/Cas9-based targeting of the promoter region.