eprintid: 10203573 rev_number: 7 eprint_status: archive userid: 699 dir: disk0/10/20/35/73 datestamp: 2025-01-20 11:19:16 lastmod: 2025-01-20 11:19:16 status_changed: 2025-01-20 11:19:16 type: article metadata_visibility: show sword_depositor: 699 creators_name: Kempthorne, Liam creators_name: Vaizoglu, Deniz creators_name: Cammack, Alexander J creators_name: Carcolé, Mireia creators_name: Roberts, Martha J creators_name: Mikheenko, Alla creators_name: Fisher, Alessia creators_name: Suklai, Pacharaporn creators_name: Muralidharan, Bhavana creators_name: Kroll, François creators_name: Moens, Thomas G creators_name: Yshii, Lidia creators_name: Verschoren, Stijn creators_name: Hölbling, Benedikt V creators_name: Moreira, Francisco C creators_name: Katona, Eszter creators_name: Coneys, Rachel creators_name: de Oliveira, Paula creators_name: Zhang, Yong-Jie creators_name: Jansen, Karen creators_name: Daughrity, Lillian M creators_name: McGown, Alexander creators_name: Ramesh, Tennore M creators_name: Van Den Bosch, Ludo creators_name: Lignani, Gabriele creators_name: Rahim, Ahad A creators_name: Coyne, Alyssa N creators_name: Petrucelli, Leonard creators_name: Rihel, Jason creators_name: Isaacs, Adrian M title: Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo ispublished: pub divisions: UCL divisions: B02 divisions: C07 divisions: C08 divisions: D07 divisions: D09 divisions: D10 divisions: F81 divisions: F85 divisions: F86 divisions: F96 divisions: G10 keywords: C9orf72 Protein, Amyotrophic Lateral Sclerosis, Humans, Frontotemporal Dementia, Animals, CRISPR-Cas Systems, RNA, Antisense, Mice, HEK293 Cells, Induced Pluripotent Stem Cells, DNA Repeat Expansion, Disease Models, Animal, Neurons, Genetic Therapy note: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. abstract: The most common genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) is an intronic G4C2 repeat expansion in C9orf72. The repeats undergo bidirectional transcription to produce sense and antisense repeat RNA species, which are translated into dipeptide repeat proteins (DPRs). As toxicity has been associated with both sense and antisense repeat-derived RNA and DPRs, targeting both strands may provide the most effective therapeutic strategy. CRISPR-Cas13 systems mature their own guide arrays, allowing targeting of multiple RNA species from a single construct. We show CRISPR-Cas13d variant CasRx effectively reduces overexpressed C9orf72 sense and antisense repeat transcripts and DPRs in HEK cells. In C9orf72 patient-derived iPSC-neuron lines, CRISPR-CasRx reduces endogenous sense and antisense repeat RNAs and DPRs and protects against glutamate-induced excitotoxicity. AAV delivery of CRISPR-CasRx to two distinct C9orf72 repeat mouse models significantly reduced both sense and antisense repeat-containing transcripts. This highlights the potential of RNA-targeting CRISPR systems as therapeutics for C9orf72 ALS/FTD. date: 2025-01-08 date_type: published publisher: Springer Science and Business Media LLC official_url: https://doi.org/10.1038/s41467-024-55550-x oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2351605 doi: 10.1038/s41467-024-55550-x medium: Electronic pii: 10.1038/s41467-024-55550-x lyricists_name: Rihel, Jason lyricists_name: Rahim, Ahad lyricists_name: Isaacs, Adrian lyricists_name: Lignani, Gabriele lyricists_name: Mikheenko, Alla lyricists_id: JRIHE66 lyricists_id: ARAHI02 lyricists_id: AISAA22 lyricists_id: GLIGN08 lyricists_id: AMIKH76 actors_name: Gaisford, Simon actors_name: Harris, Jean actors_id: SGAIS88 actors_id: JAHAR68 actors_role: owner actors_role: impersonator funding_acknowledgements: 648716 - C9ND [EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council)]; 217150/Z/19/Z [Wellcome Trust (Wellcome)] full_text_status: public publication: Nature Communications volume: 16 article_number: 459 event_location: England issn: 2041-1723 citation: Kempthorne, Liam; Vaizoglu, Deniz; Cammack, Alexander J; Carcolé, Mireia; Roberts, Martha J; Mikheenko, Alla; Fisher, Alessia; ... Isaacs, Adrian M; + view all <#> Kempthorne, Liam; Vaizoglu, Deniz; Cammack, Alexander J; Carcolé, Mireia; Roberts, Martha J; Mikheenko, Alla; Fisher, Alessia; Suklai, Pacharaporn; Muralidharan, Bhavana; Kroll, François; Moens, Thomas G; Yshii, Lidia; Verschoren, Stijn; Hölbling, Benedikt V; Moreira, Francisco C; Katona, Eszter; Coneys, Rachel; de Oliveira, Paula; Zhang, Yong-Jie; Jansen, Karen; Daughrity, Lillian M; McGown, Alexander; Ramesh, Tennore M; Van Den Bosch, Ludo; Lignani, Gabriele; Rahim, Ahad A; Coyne, Alyssa N; Petrucelli, Leonard; Rihel, Jason; Isaacs, Adrian M; - view fewer <#> (2025) Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo. Nature Communications , 16 , Article 459. 10.1038/s41467-024-55550-x <https://doi.org/10.1038/s41467-024-55550-x>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10203573/1/s41467-024-55550-x.pdf