The sustained expression of Cas9 targeting toxic RNAs reverses disease phenotypes in mouse models of myotonic dystrophy type 1

Batra, Ranjan; Nelles, David A.; Roth, Daniela; Krach, Florian; Nutter, Curtis A.; Tadokoro, Takahiro; Thomas, James; Sznajder, Łukasz J.; Blue, Steven M.; Gutierrez, Haydee L.; Liu, Patrick; Aigner, Stefan; Platoshyn, Oleksandr; Miyanohara, Atsushi; Maršala, Martin; Swanson, Maurice S.; Yeo, G

doi:10.1038/s41551-020-00607-7

Cited by 41 publications

(42 citation statements)

References 83 publications

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“…In a recent demonstration from our group, we observed that sustained expression of virally-encoded rCas9-PIN targeting expanded CAG repeat RNA reversed several phenotypes in neonatal and adult mouse models of DM1 ( Batra et al 2020 ). Our group noted that intramuscular or systemic injections of AAV-encoding rCas9-PIN with CAG-targeting sgRNA resulted in expression that endured for nearly 3 months with sustained specificity for expanded CAG repeats.…”

Section: Reductions Of Toxic Rna Levelsmentioning

confidence: 85%

“…Our group noted that intramuscular or systemic injections of AAV-encoding rCas9-PIN with CAG-targeting sgRNA resulted in expression that endured for nearly 3 months with sustained specificity for expanded CAG repeats. While future experiments will seek to more thoroughly characterize in vivo phenotypes, identify off-target effects, and mitigate the inherent immunogenicity from expression of non-self Cas9 fusions, this work highlights the feasibility of prolonged AAV-mediated expression of rCas9-PIN for targeting a variety of MRE disorders ( Batra et al 2020 ).…”

Section: Reductions Of Toxic Rna Levelsmentioning

confidence: 99%

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Repeat RNA expansion disorders of the nervous system: post-transcriptional mechanisms and therapeutic strategies

Schwartz

Jones

Yeo

2020

Critical Reviews in Biochemistry and Molecular Biology

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Dozens of incurable neurological disorders result from expansion of short repeat sequences in both coding and non-coding regions of the transcriptome. Short repeat expansions underlie microsatellite repeat expansion (MRE) disorders including myotonic dystrophy (DM1, CUG 50–3,500 in DMPK ; DM2, CCTG 75–11,000 in ZNF9), fragile X tremor ataxia syndrome (FXTAS, CGG 50–200 in FMR1), spinal bulbar muscular atrophy (SBMA, CAG 40–55 in AR), Huntington’s disease (HD, CAG 36–121 in HTT), C9ORF72-amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD and C9-ALS/FTD, GGGGCC in C9ORF72), and many others, like ataxias. Recent research has highlighted several mechanisms that may contribute to pathology in this heterogeneous class of neurological MRE disorders – bidirectional transcription, intranuclear RNA foci, and repeat associated non-AUG (RAN) translation – which are the subject of this review. Additionally, many MRE disorders share similar underlying molecular pathologies that have been recently targeted in experimental and preclinical contexts. We discuss the therapeutic potential of versatile therapeutic strategies that may selectively target disrupted RNA-based processes and may be readily adaptable for the treatment of multiple MRE disorders. Collectively, the strategies under consideration for treatment of multiple MRE disorders include reducing levels of toxic RNA, preventing RNA foci formation, and eliminating the downstream cellular toxicity associated with peptide repeats produced by RAN translation. While treatments are still lacking for the majority of MRE disorders, several promising therapeutic strategies have emerged and will be evaluated within this review.

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Section: Reductions Of Toxic Rna Levelsmentioning

confidence: 85%

Section: Reductions Of Toxic Rna Levelsmentioning

confidence: 99%

Repeat RNA expansion disorders of the nervous system: post-transcriptional mechanisms and therapeutic strategies

Schwartz

Jones

Yeo

2020

Critical Reviews in Biochemistry and Molecular Biology

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“…DM1 expansions induce formation of nuclear RNA foci that can be visually detected by fluorescent in situ hybridization (RNA FISH) 69 . We analysed data from two studies with RNA-seq of cells treated with CUG-degrading dCas9 70,71 . The first study was performed in myotubes derived from induced pluripotent stem cells from a DM1 patient and an unaffected individual 70 .…”

Section: Myotonic Dystrophy 1 and Long Ctg Repeat Cell Line Experimentsmentioning

confidence: 99%

“…RNA-seq data for SCA3 was obtained from a study describing the transcriptome of peripheral blood mononuclear cells in 12 SCA3-affected individuals with 12 age-and gender-matched 71 . This study used Illumina TruSeq PolyA library preparation, and samples were sequenced on an Illumina HiSeq 4000 instrument.…”

Section: Transcriptome Sequence Datamentioning

confidence: 99%

Ultrafast, alignment-free detection of repeat expansions in next-generation DNA and RNA sequencing data

Fearnley

Bennett

Bahlo

2021

Preprint

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Short tandem repeat expansions are an established cause of diseases such as Huntington's disease. Bioinformatic methods for detecting repeat expansions in short-read sequencing have revealed new repeat expansions in humans. Current bioinformatic methods to detect repeat expansions require alignment information to identify repetitive motif enrichment at genomic locations. We present superSTR, an ultrafast method that does not require alignment, capable of efficiently processing DNA and RNA sequencing data. superSTR is applied to UK Biobank data to efficiently analyse 49,953 whole exomes in a screening experiment, identifying known mutations as well as diseases not previously associated with REs. superSTR also identifies repeat expansion motifs in RNAseq data, demonstrated in several disorders and cell lines. superSTR is a new tool for the most efficient repeat expansion detection currently possible and complements existing locus-specific, reference dependent repeat expansion analysis tools. superSTR is available from https://github.com/bahlolab/superSTR.

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“…Reporting in Nature Biomedical Engineering, Gene Yeo and colleagues now show the performance of such a RNA-targeting strategy in a well-characterized mouse model of DM1 (ref. 13 ): human skeletal actin long-repeat (HSA LR ) transgenic mice harbouring 220 CTG repeats located within the 3′ untranslated region of the human gene for actin 14 . HSA LR mice express expanded CUG repeats exclusively in skeletal muscles, and display DM1-associated phenotypes, including nuclear RNA foci, alternative-splicing dysregulations, muscle defects and myotonia.…”

Section: Denis Furlingmentioning

confidence: 99%

Cas9 targeting of toxic foci of RNA repeats

Furling

2021

Nat Biomed Eng

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The sustained expression of Cas9 targeting toxic RNAs reverses disease phenotypes in mouse models of myotonic dystrophy type 1

Cited by 41 publications

References 83 publications

Repeat RNA expansion disorders of the nervous system: post-transcriptional mechanisms and therapeutic strategies

Repeat RNA expansion disorders of the nervous system: post-transcriptional mechanisms and therapeutic strategies

Ultrafast, alignment-free detection of repeat expansions in next-generation DNA and RNA sequencing data

Cas9 targeting of toxic foci of RNA repeats

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