TY - JOUR
T1 - A high-fidelity CRISPR-Cas13 system improves abnormalities associated with C9ORF72-linked ALS/FTD
AU - McCallister, Tristan X.
AU - Lim, Colin K.W.
AU - Singh, Mayuri
AU - Zhang, Sijia
AU - Ahsan, Najah S.
AU - Terpstra, William M.
AU - Xiong, Alisha Y.
AU - Zeballos C, M. Alejandra
AU - Powell, Jackson E.
AU - Drnevich, Jenny
AU - Kang, Yifei
AU - Gaj, Thomas
N1 - We thank A. Hernandez for helpful discussion and assistance with library preparations for NGS and P. Perez-Pinera for access to the Synergy HTX Plate Reader. This work was supported by the NIH/NINDS (1U01NS122102-01A1 and 1R01NS123556-01A1 to T.G.), the NIH/NIGMS (5R01GM141296 to T.G.), the Muscular Dystrophy Association (MDA602798 to T.G.), the Simons Foundation (887187 to T.G.), the Parkinson\u2019s Disease Foundation (PF-IMP-1950 to T.G.), the Judith & Jean Pape Adams Foundation and the ALS Association (20-IIP-516 to T.G.). M.A.Z.C. was supported by the NIH/NIBIB (T32EB019944), the Mavis Future Faculty Fellows Program, and an Aspire Fellowship from the University of Illinois Urbana-Champaign. Cartoons were created with BioRender.
PY - 2025/12
Y1 - 2025/12
N2 - An abnormal expansion of a GGGGCC (G4C2) hexanucleotide repeat in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two debilitating neurodegenerative disorders driven in part by gain-of-function mechanisms involving transcribed forms of the repeat expansion. By utilizing a Cas13 variant with reduced collateral effects, we develop here a high-fidelity RNA-targeting CRISPR-based system for C9ORF72-linked ALS/FTD. When delivered to the brain of a transgenic rodent model, this Cas13-based platform curbed the expression of the G4C2 repeat-containing RNA without affecting normal C9ORF72 levels, which in turn decreased the formation of RNA foci, reduced the production of a dipeptide repeat protein, and reversed transcriptional deficits. This high-fidelity system possessed improved transcriptome-wide specificity compared to its native form and mediated targeting in motor neuron-like cells derived from a patient with ALS. These results lay the foundation for the implementation of RNA-targeting CRISPR technologies for C9ORF72-linked ALS/FTD.
AB - An abnormal expansion of a GGGGCC (G4C2) hexanucleotide repeat in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two debilitating neurodegenerative disorders driven in part by gain-of-function mechanisms involving transcribed forms of the repeat expansion. By utilizing a Cas13 variant with reduced collateral effects, we develop here a high-fidelity RNA-targeting CRISPR-based system for C9ORF72-linked ALS/FTD. When delivered to the brain of a transgenic rodent model, this Cas13-based platform curbed the expression of the G4C2 repeat-containing RNA without affecting normal C9ORF72 levels, which in turn decreased the formation of RNA foci, reduced the production of a dipeptide repeat protein, and reversed transcriptional deficits. This high-fidelity system possessed improved transcriptome-wide specificity compared to its native form and mediated targeting in motor neuron-like cells derived from a patient with ALS. These results lay the foundation for the implementation of RNA-targeting CRISPR technologies for C9ORF72-linked ALS/FTD.
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U2 - 10.1038/s41467-024-55548-5
DO - 10.1038/s41467-024-55548-5
M3 - Article
C2 - 39779681
AN - SCOPUS:85215129696
SN - 2041-1723
VL - 16
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 460
ER -