Cis -splicing and Translation of the Pre- Trans -splicing Molecule Combine With Efficiency in Spliceosome-mediated RNA Trans -splicing

Monjaret, François; Bourg, Nathalie; Suel, Laurence; Roudaut, Carinne; Roy, Florence Le; Richard, Isabelle; Charton, Karine

doi:10.1038/mt.2014.35

Cited by 32 publications

(26 citation statements)

References 32 publications

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“…In 2012, Rindt et al demonstrated in cellular models of Huntington's disease that the mutated exon can be eliminated by 5′ replacement from the Huntingtin encoding mRNA . Recently, Monjaret et al described trans ‐splicing of the Titin pre‐mRNA, whose mutations lead to muscular dystrophies . In this study, the trans ‐spliced product was not accurately quantified and appeared clearly minor.…”

Section: Principle Advantages and Applications Of Spliceosome‐mediamentioning

confidence: 71%

“…In the case of a recessive disease, it is likely that correcting only part of the mRNA will result in therapeutic benefit, because one ends up in the scenario of a heterozygous individual who presents no phenotype. SMaRT technology has thus been evaluated for a significant number of recessive pathologies, such as cystic fibrosis, hemophilia A, dysferlinopathies and titinopathies, X‐linked immunodeficiency with hyper‐IgM, spinal muscular atrophy, severe combined immune deficiency, Duchenne muscular dystrophy, and epidermolysis bullosa . All these studies have shown that it is possible to partially correct the cellular pool of mutated mRNA.…”

Section: Principle Advantages and Applications Of Spliceosome‐mediamentioning

confidence: 99%

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mRNAtrans‐splicing in gene therapy for genetic diseases

et al. 2016

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Spliceosome‐mediated RNA trans‐splicing, or SMaRT, is a promising strategy to design innovative gene therapy solutions for currently intractable genetic diseases. SMaRT relies on the correction of mutations at the post‐transcriptional level by modifying the mRNA sequence. To achieve this, an exogenous RNA is introduced into the target cell, usually by means of gene transfer, to induce a splice event in trans between the exogenous RNA and the target endogenous pre‐mRNA. This produces a chimeric mRNA composed partly of exons of the latter, and partly of exons of the former, encoding a sequence free of mutations. The principal challenge of SMaRT technology is to achieve a reaction as complete as possible, i.e., resulting in 100% repairing of the endogenous mRNA target. The proof of concept of SMaRT feasibility has already been established in several models of genetic diseases caused by recessive mutations. In such cases, in fact, the repair of only a portion of the mutant mRNA pool may be sufficient to obtain a significant therapeutic effect. However in the case of dominant mutations, the target cell must be freed from the majority of mutant mRNA copies, requiring a highly efficient trans‐splicing reaction. This likely explains why only a few examples of SMaRT approaches targeting dominant mutations are reported in the literature. In this review, we explain in details the mechanism of trans‐splicing, review the different strategies that are under evaluation to lead to efficient trans‐splicing, and discuss the advantages and limitations of SMaRT. WIREs RNA 2016, 7:487–498. doi: 10.1002/wrna.1347For further resources related to this article, please visit the WIREs website.

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Section: Principle Advantages and Applications Of Spliceosome‐mediamentioning

confidence: 71%

Section: Principle Advantages and Applications Of Spliceosome‐mediamentioning

confidence: 99%

mRNAtrans‐splicing in gene therapy for genetic diseases

et al. 2016

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“…Although RNA trans -splicing-induced RNA repair strategies were successfully applied for the restoration of gene functions in preclinical models for various diseases such as cystic fibrosis, 25, 26, 27 spinal muscular atrophy, 28, 29, 30 haemophilia A, 31 tauopathies, 32, 33 EB simplex, 34 X-linked immunodeficiency with hyper-IgM, 35 severe combined immune deficiency, 36 myotonic dystrophy type 1, 37 frontotemporal dementia with parkinsonism linked to chromosome 17, 32 Duchenne muscular dystrophy 38 and dysferlinopathy, 39 there is still scope for improvement of the trans -splicing efficiency to achieve a sustained and efficient correction of the mutated transcript. In recent studies, we have reported that the BD of the RTM is crucial for the efficiency of the trans -splicing process.…”

Section: Discussionmentioning

confidence: 99%

Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations

et al. 2016

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RNA trans-splicing has become a versatile tool in the gene therapy of monogenetic diseases. This technique is especially valuable for the correction of mutations in large genes such as COL7A1, which underlie the dystrophic subtype of the skin blistering disease epidermolysis bullosa. Over 800 mutations spanning the entire length of the COL7A1 gene have been associated with defects in type VII collagen, leading to excessive fragility of epithelial tissues, the hallmark of dystrophic epidermolysis bullosa (DEB). In the present study, we designed an RNA trans-splicing molecule (RTM) that is capable of repairing any given mutation within a 4200 nucleotide region spanning the 3′ half of COL7A1. The selected RTM, RTM28, was able to induce accurate trans-splicing into endogenous COL7A1 pre-mRNA transcripts in a type VII collagen-deficient DEB patient-derived cell line. Correct trans-splicing was detected at the RNA level by semiquantitative RT-PCR and correction of full-length type VII collagen was confirmed at the protein level by immunofluorescence and western blot analyses. Our results demonstrate that RTM28, which covers >60% of all mutations reported in DEB and is thus the longest RTM described so far for the repair of COL7A1, represents a promising candidate for therapeutic applications.

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“…Approaches based on 3′-replacement trans- splicing have already been used, both in vitro and in vivo , to correct several genetic diseases, including cystic fibrosis, 27 , 28 , 29 spinal muscular atrophy, 30 , 31 , 32 hemophilia A, 33 tauopathies, 34 , 35 epidermolysis bullosa simplex, 36 X-linked immunodeficiency with hyper-IgM, 37 severe combined immune deficiency, 38 myotonic dystrophy type 1, 39 frontotemporal dementia with parkinsonism linked to chromosome 17, 34 Duchenne muscular dystrophy, 40 dysferlinopathy. 41 Trans- splicing has also recently been considered as a possible basis of new strategies for treating cancer, 42 , 43 producing therapeutic proteins 44 and for molecular imaging. 45 …”

Section: Introductionmentioning

confidence: 99%

Repair of Rhodopsin mRNA by Spliceosome-Mediated RNA Trans -Splicing: A New Approach for Autosomal Dominant Retinitis Pigmentosa

et al. 2015

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The promising clinical results obtained for ocular gene therapy in recent years have paved the way for gene supplementation to treat recessively inherited forms of retinal degeneration. The situation is more complex for dominant mutations, as the toxic mutant gene product must be removed. We used spliceosome-mediated RNA trans-splicing as a strategy for repairing the transcript of the rhodopsin gene, the gene most frequently mutated in autosomal dominant retinitis pigmentosa. We tested 17 different molecules targeting the pre-mRNA intron 1, by transient transfection of HEK-293T cells, with subsequent trans-splicing quantification at the transcript level. We found that the targeting of some parts of the intron promoted trans-splicing more efficiently than the targeting of other areas, and that trans-splicing rate could be increased by modifying the replacement sequence. We then developed cell lines stably expressing the rhodopsin gene, for the assessment of phenotypic criteria relevant to the pathogenesis of retinitis pigmentosa. Using this model, we showed that trans-splicing restored the correct localization of the protein to the plasma membrane. Finally, we tested our best candidate by AAV gene transfer in a mouse model of retinitis pigmentosa that expresses a mutant allele of the human rhodopsin gene, and demonstrated the feasibility of trans-splicing in vivo. This work paves the way for trans-splicing gene therapy to treat retinitis pigmentosa due to rhodopsin gene mutation and, more generally, for the treatment of genetic diseases with dominant transmission.

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Cis -splicing and Translation of the Pre- Trans -splicing Molecule Combine With Efficiency in Spliceosome-mediated RNA Trans -splicing

Cited by 32 publications

References 32 publications

mRNAtrans‐splicing in gene therapy for genetic diseases

mRNAtrans‐splicing in gene therapy for genetic diseases

Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations

Repair of Rhodopsin mRNA by Spliceosome-Mediated RNA Trans -Splicing: A New Approach for Autosomal Dominant Retinitis Pigmentosa

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