Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo
Abstract
AbstractThe 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.
Article Details
Authors (30)
Liam Kempthorne
Deniz Vaizoglu
Alexander J. Cammack
Mireia Carcolé
Martha J. Roberts
Alla Mikheenko
Alessia Fisher
Pacharaporn Suklai
Bhavana Muralidharan
François Kroll
Thomas G. Moens
Lidia Yshii
Stijn Verschoren
Benedikt V. Hölbling
Francisco C. Moreira
Eszter Katona
Rachel Coneys
Paula de Oliveira
Yong-Jie Zhang
Karen Jansen
Lillian M. Daughrity
Alexander McGown
Tennore M. Ramesh
Ludo Van Den Bosch
Gabriele Lignani
Research Department of Epilepsy, Queen Square Institute of Neurology, University College London, London, UK.
Ahad A. Rahim
Department of Pharmacology, University College London School of Pharmacy, University College London
Alyssa N. Coyne
Leonard Petrucelli
Department of Neuroscience, Mayo Clinic
Jason Rihel
Adrian M. Isaacs