Dystrophin gene mutations are the cause of dystrophinopathies, a group of phenotypes manifesting with striated muscle involvement. The pathogenesis of dystrophinopathies as well as the genotype-phenotype correlation have not been entirely elucidated. The project proposed a strategy to identify, characterise and modulate non–coding regulatory sequences, with particular attention to the splicing regulatory motifs. The approach takes advantage of a large series of patients with dystrophinopathies already characterised. Among these, we selected two categories: 1) in which the causative mutation is disobeying the Monaco frame theory in terms of phenotype/genotype relationship, 2) in which no mutations at the coding level have been found. In these selected cases, we will analyse both the genomic configuration and the RNA editing in the available tissues in order to obtain the breakpoint definition and to identify mutations located within non-coding regions. We will also focalise our attention on LINE1 mobile elements in the dystrophin gene, given their role in the human genome and their involvement in genetic diseases. We will evaluate both their structural/functional characteristics and their relationship with dystrophin mutations by exploring their transcription status and the occurrence in them of functional motifs. Following these steps, sequence comparative analysis will focus the presence within the intronic regions explored of conserved non-coding sequences, and functional assays will allow the evaluation of their role in splicing modulation. Antisense interference will allow both the confirmation of the functional significance of the identified motifs in splicing and attempt a modulation of the splicing process via a restoration of the wild type RNA phenotype. This approach has been shown to be a promising tool for gene expression interference and might have possible therapeutic implications

A strategy to identify and modulate sequences-motifs relevant for the pathogenesis of dystrophinopathies by functional characterisation of non-coding regions involved in dystrophin gene expression regulation (Telethon GGPO2112)

FERLINI, Alessandra
2002

Abstract

Dystrophin gene mutations are the cause of dystrophinopathies, a group of phenotypes manifesting with striated muscle involvement. The pathogenesis of dystrophinopathies as well as the genotype-phenotype correlation have not been entirely elucidated. The project proposed a strategy to identify, characterise and modulate non–coding regulatory sequences, with particular attention to the splicing regulatory motifs. The approach takes advantage of a large series of patients with dystrophinopathies already characterised. Among these, we selected two categories: 1) in which the causative mutation is disobeying the Monaco frame theory in terms of phenotype/genotype relationship, 2) in which no mutations at the coding level have been found. In these selected cases, we will analyse both the genomic configuration and the RNA editing in the available tissues in order to obtain the breakpoint definition and to identify mutations located within non-coding regions. We will also focalise our attention on LINE1 mobile elements in the dystrophin gene, given their role in the human genome and their involvement in genetic diseases. We will evaluate both their structural/functional characteristics and their relationship with dystrophin mutations by exploring their transcription status and the occurrence in them of functional motifs. Following these steps, sequence comparative analysis will focus the presence within the intronic regions explored of conserved non-coding sequences, and functional assays will allow the evaluation of their role in splicing modulation. Antisense interference will allow both the confirmation of the functional significance of the identified motifs in splicing and attempt a modulation of the splicing process via a restoration of the wild type RNA phenotype. This approach has been shown to be a promising tool for gene expression interference and might have possible therapeutic implications
Ferlini, Alessandra
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/1679298
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