A Bump-Hole Approach for Directed RNA Editing

Monteleone, Leanna R.; Matthews, Melissa M.; Palumbo, Cody M.; Thomas, Justin M.; Zheng, Yuxuan; Chiang, Yang‐Jen; Fisher, A.J.; Beal, Peter A.

doi:10.1016/j.chembiol.2018.10.025

Cited by 29 publications

(34 citation statements)

References 55 publications

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“…An advantage of this finding is that future experiments to evaluate rescue of Rett syndrome-like phenotypes in mice will result in lower levels of editase per cell because of peripheral virus delivery, which is necessary to infect the entire brain. Additionally, several groups are already beginning to identify editase molecules with higher specificity and efficiency ( Cox et al, 2017 ; Monteleone et al, 2019 ) for in vivo testing.…”

Section: Discussionmentioning

confidence: 99%

In Vivo Repair of a Protein Underlying a Neurological Disorder by Programmable RNA Editing

et al. 2020

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SUMMARY Programmable RNA editing is gaining momentum as an approach to repair mutations, but its efficiency in repairing endogenous mutant RNA in complex tissue is unknown. Here we apply this approach to the brain and successfully repair a guanosine-to-adenosine mutation in methyl CpG binding protein 2 RNA that causes the neurodevelopmental disease Rett syndrome. Repair is mediated by hippocampal injections of juvenile Mecp2 317G>A mice with an adeno-associated virus expressing the hyperactive catalytic domain of adenosine deaminase acting on RNA 2 and Mecp2 guide. After 1 month, 50% of Mecp2 RNA is recoded in three different hippocampal neuronal populations. MeCP2 protein localization to heterochromatin is restored in neurons to 50% of wild-type levels. Whole-transcriptome RNA analysis of one neuronal population indicates that the majority of off-target editing sites exhibit rates of 30% or less. This study demonstrates that programmable RNA editing can be utilized to repair mutations in mouse models of neurological disease.

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Section: Discussionmentioning

confidence: 99%

In Vivo Repair of a Protein Underlying a Neurological Disorder by Programmable RNA Editing

et al. 2020

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“…However, these mutant ADAR versions could lead to significant off-target effects. To mitigate this risk, Monteleone et al showed that they were able to reduce the off-target effects by mutating ADAR2 and altering the guide RNA [ 81 ]. Vallecillo-Viejo et al also showed that the off-target effects could be reduced when artificial versions of ADAR were targeted to the nucleus (through attaching several NLS to the artificial ADAR) [ 82 ].…”

Section: Suppression Of Nonsense Mutations: Approaches and Challenmentioning

confidence: 99%

Suppression of Nonsense Mutations by New Emerging Technologies

Morais

Adachi

2020

IJMS

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Nonsense mutations often result from single nucleotide substitutions that change a sense codon (coding for an amino acid) to a nonsense or premature termination codon (PTC) within the coding region of a gene. The impact of nonsense mutations is two-fold: (1) the PTC-containing mRNA is degraded by a surveillance pathway called nonsense-mediated mRNA decay (NMD) and (2) protein translation stops prematurely at the PTC codon, and thus no functional full-length protein is produced. As such, nonsense mutations result in a large number of human diseases. Nonsense suppression is a strategy that aims to correct the defects of hundreds of genetic disorders and reverse disease phenotypes and conditions. While most clinical trials have been performed with small molecules, there is an increasing need for sequence-specific repair approaches that are safer and adaptable to personalized medicine. Here, we discuss recent advances in both conventional strategies as well as new technologies. Several of these will soon be tested in clinical trials as nonsense therapies, even if they still have some limitations and challenges to overcome.

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“…Our case is likely the worst case scenario because we injected directly into hippocampus a very high titer of virus, and systemic injections, required for behavioral analysis in Rett mouse models, will result in much lower expression of Editase/cell. Several groups are already beginning to identify Editase molecules with higher specificity and efficiency [17,49], which will need to be tested in vivo, and the Rett mouse models will again be ideal for determining phenotypes. Importantly, despite the off-target events, in the mice expressing Editase and Mecp2 targeting guide, we observed mean editing rates of 50% at the target adenosines in Mecp2 RNA, in three different hippocampal neuronal fields containing multiple neuronal types [50][51][52][53][54] (Figure 2b).…”

Section: Discussionmentioning

confidence: 99%

In vivo repair of a protein underlying a neurological disorder by programmable RNA editing

Sinnamon

Kim

Fisk

et al. 2020

Preprint

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RNA base editing is gaining momentum as an approach to repair mutations, but its application to neurological disease has not been established. We have succeeded in directed transcript editing of a pathological mutation in a mouse model of the neurodevelopmental disease, Rett syndrome. Specifically, we directed editing of a guanosine to adenosine mutation in RNA encoding Methyl CpG Binding Protein 2 (MECP2). Repair was mediated by injecting the hippocampus of juvenile Rett mice with an adeno-associated virus expressing both an engineered enzyme containing the catalytic domain of Adenosine Deaminase Acting on RNA 2 and a Mecp2 targeting guide. After one month, 50% of Mecp2 RNA was recoded in three different hippocampal neuronal subtypes, and the ability of MeCP2 protein to associate with heterochromatin was similarly restored to 50% of wild-type levels. This study represents the first in vivo programmable RNA editing applied to a model of neurological disease.

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A Bump-Hole Approach for Directed RNA Editing

Cited by 29 publications

References 55 publications

In Vivo Repair of a Protein Underlying a Neurological Disorder by Programmable RNA Editing

In Vivo Repair of a Protein Underlying a Neurological Disorder by Programmable RNA Editing

Suppression of Nonsense Mutations by New Emerging Technologies

In vivo repair of a protein underlying a neurological disorder by programmable RNA editing

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