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article cooper-knock chromosome 9p21-linked als-ftd large scale view amyotrophic lateral sclerosis article download pdf chromosome 9p-linked ftd formalin fixed paraffin-embedded aggregating dipeptide-repeat proteins uv-crosslinking studies confirmed likelihood-ratio test revealed depth clinico-pathological examination ipsc-derived motor neurons interacting rna-processing factors dipeptide repeat protein post-mitotic mature cells represents end-stage disease rna fish probes als/ftd c9orf72 expansion repeat rna sequences uv-crosslinking studies showed cell-specific transcriptional regulation secondary structures mackenzie ir antisense/sense repeat sequence hexanucleotide repeat expansion dipeptide repeat proteins rna-binding activity national research committee privacy choices/manage cookies conducted extensive immunohistochemistry donnelly cj c9orf72 repeat expansions ftd cases including antisense repeat rnas c9orf72-related disease original author intra molecular interactions medical research council protein binding partners full access myotonic dystrophy type rna-binding factors rna recognition motif c9orf72 repeat expansion c9orf72 ggggcc repeat antisense repeat rna author information authors uv-crosslinking studies clinico-pathological correlations common genetic variant

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         description:GGGGCC repeat expansions of C9ORF72 represent the most common genetic variant of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. We and others have proposed that RNA transcribed from the repeat sequence is toxic via sequestration of RNA-binding factors. Both GGGGCC-repeat (sense) and CCCCGG-repeat (antisense) molecules are detectable by fluorescence in situ hybridisation as RNA foci, but their relative expression pattern within the CNS and contribution to disease has not been determined. Blinded examination of CNS biosamples from ALS patients with a repeat expansion of C9ORF72 showed that antisense foci are present at a significantly higher frequency in cerebellar Purkinje neurons and motor neurons, whereas sense foci are present at a significantly higher frequency in cerebellar granule neurons. Consistent with this, inclusions containing sense or antisense derived dipeptide repeat proteins were present at significantly higher frequency in cerebellar granule neurons or motor neurons, respectively. Immunohistochemistry and UV-crosslinking studies showed that sense and antisense RNA molecules share similar interactions with SRSF2, hnRNP K, hnRNP A1, ALYREF, and hnRNP H/F. Together these data suggest that, although sense and antisense RNA molecules might be expected to be equally toxic via their shared protein binding partners, distinct patterns of expression in various CNS neuronal populations could lead to relative differences in their contribution to the pathogenesis of neuronal injury. Moreover in motor neurons, which are the primary target of pathology in ALS, the presence of antisense foci (χ 2, p < 0.00001) but not sense foci (χ 2, p = 0.75) correlated with mislocalisation of TDP-43, which is the hallmark of ALS neurodegeneration. This has implications for translational approaches to C9ORF72 disease, and furthermore interacting RNA-processing factors and transcriptional activators responsible for antisense versus sense transcription might represent novel therapeutic targets.
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         Amyotrophic lateral sclerosis
         RNA foci
         Dipeptide repeat protein
         Immunohistochemistry
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