Mapping the self-association domains of ataxin-1: Identification of novel non overlapping motifs

Aggregation Process ◽

Combined Approach ◽

Polyglutamine Tract ◽

Self Association

The neurodegenerative spinocerebellar ataxia type 1 (SCA1) is caused by aggregation and misfolding of the ataxin-1 protein. While the pathology correlates with mutations that lead to expansion of a polyglutamine tract in the protein, other regions contribute to the aggregation process as also non-expanded ataxin-1 is intrinsically aggregation-prone and forms nuclear foci in cell. Here, we have used a combined approach based on FRET analysis, confocal microscopy and in vitro techniques to map aggregation-prone regions other than polyglutamine and to establish the importance of dimerization in self-association/foci formation. Identification of aggregation-prone regions other than polyglutamine could greatly help the development of SCA1 treatment more specific than that based on targeting the low complexity polyglutamine region.

Mapping the self-association domains of ataxin-1: Identification of novel non overlapping motifs

10.7287/peerj.preprints.259 ◽

2014 ◽

Author(s):

Rajesh Menon ◽

Daniel Soong ◽

Cesira de Chiara ◽

Mark Holt ◽

John McCormick ◽

...

Keyword(s):

Low Complexity ◽

The Self ◽

Aggregation Process ◽

Combined Approach ◽

Polyglutamine Tract ◽

Self Association

The neurodegenerative spinocerebellar ataxia type 1 (SCA1) is caused by aggregation and misfolding of the ataxin-1 protein. While the pathology correlates with mutations that lead to expansion of a polyglutamine tract in the protein, other regions contribute to the aggregation process as also non-expanded ataxin-1 is intrinsically aggregation-prone and forms nuclear foci in cell. Here, we have used a combined approach based on FRET analysis, confocal microscopy and in vitro techniques to map aggregation-prone regions other than polyglutamine and to establish the importance of dimerization in self-association/foci formation. Identification of aggregation-prone regions other than polyglutamine could greatly help the development of SCA1 treatment more specific than that based on targeting the low complexity polyglutamine region.

The spinocerebellar ataxia type 1 protein, ataxin-1, has RNA-binding activity that is inversely affected by the length of its polyglutamine tract

Human Molecular Genetics ◽

10.1093/hmg/10.1.25 ◽

2001 ◽

Vol 10 (1) ◽

pp. 25-30 ◽

Cited By ~ 105

Author(s):

S. Yue

Keyword(s):

Spinocerebellar Ataxia ◽

Rna Binding ◽

Binding Activity ◽

Polyglutamine Tract

CRISPR/Cas9-based silencing of the ATXN1 gene in Spinocerebellar ataxia type 1 (SCA1) fibroblasts

10.1101/2020.07.04.187559 ◽

2020 ◽

Author(s):

Francesca Salvatori ◽

Mariangela Pappadà ◽

Mariaconcetta Sicurella ◽

Mattia Buratto ◽

Valentina Simioni ◽

...

Keyword(s):

Spinocerebellar Ataxia ◽

Neurodegenerative Disorder ◽

Cag Repeats ◽

Rna Seq ◽

Coding Region ◽

Polyq Tract

AbstractSpinocerebellar Ataxia type 1 (SCA1) is an autosomal dominant neurodegenerative disorder caused by a gain-of-function protein with toxic activities, containing an expanded polyQ tract in the coding region. Actually, there are no treatments available to delay the onset, stop or slow down the progression of this pathology. Many approaches developed over the years involve the use of siRNAs and antisense oligonucleotides (ASOs). Here we develop and validate a CRISPR/Cas9 therapeutic strategy in fibroblasts isolated from SCA1 patients. We started from the screening of 10 different sgRNAs able to recognize regions upstream and downstream the CAG repeats, in exon 8 of ATXN1 gene. The two most promising sgRNAs, G3 and G8, whose efficiency was evaluated with an in vitro system, significantly downregulated the ATXN 1 protein expression. This downregulation was due to the introduction of indels mutations into the ATXN1 gene. Notably, with an RNA-seq analysis, we demonstrated minimal off-target effects of our sgRNAs. These preliminary results support CRISPR/Cas9 as a promising approach for treated polyQ-expanded diseases.