Department of Biotechnology
inStem (Institute for Stem Cell Science and Regenerative Medicine)

Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo.

Publication Type

Journal Article

Date of Publication

January 8, 2025

Journal

Nature communications

Volume/Issue

16/1

ISSN

2041-1723

The most common genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) is an intronic GC 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.

Alternate Journal

Nat Commun

PubMed ID

39779704

PubMed Central ID

PMC11711508

Authors

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
Ahad A Rahim
Alyssa N Coyne
Leonard Petrucelli
Jason Rihel
Adrian M Isaacs

Keywords

Amyotrophic Lateral Sclerosis
Frontotemporal Dementia
CRISPR-Cas Systems
RNA, Antisense
HEK293 Cells
Induced Pluripotent Stem Cells
Animals
DNA Repeat Expansion
Disease Models, Animal
Mice
Neurons
Humans
Genetic Therapy
C9orf72 Protein