GSE103997 Impeding transcription of expanded microsatellite repeats by deactivated Cas9

Contributors : Belinda S Pinto ; Tanvi Saxena ; Ruan Oliveira ; Hector R Mendez-Gomez ; John D Cleary ; Lance T Denes ; Ona McConnell ; Juan Arboleda ; Guangbin Xia ; Maurice Swanson ; Eric T WangSeries Type : Expression profiling by high throughput sequencingOrganism : Homo sapiensTranscription of expanded microsatellite repeats is associated with multiple human diseases, including myotonic dystrophy, Fuchs ’ endothelial corneal dystrophy, and C9orf72-ALS/FTD. Eliminating or reducing production of RNA and proteins arising from these expanded loci holds therapeutic benefit. Here, we tested the hypothesis that a deactivated form of the Cas9 enzyme impedes transcription across expanded microsatellites. We observed a repeat length-, PAM-, and strand-dependent reduction in the abundance of repeat-containing RNAs upon targeting dCas9 directly to repeat sequences. Aberrant splicing patterns were rescued in DM1 cells, and production of RAN peptides characteristic of DM1, DM2, and C9orf72-ALS/FTD cells was drastically decreased. Systemic delivery of dCas9/gRNA by adeno-associated virus led to reductions in pathological RNA foci, rescue of chloride channel 1 protein expression, and decreased myotonia. These observations suggest that transcription of microsatellite repeat-containing RNAs is more sen sitive to perturbation than transcription of other RNAs, indicating potentially viable strategies for therapeutic intervention.
Source: GEO: Gene Expression Omnibus - Category: Genetics & Stem Cells Tags: Expression profiling by high throughput sequencing Homo sapiens Source Type: research