Identification of allele-specific RNAi effectors targeting genetic forms of Parkinson's disease.

Title: Identification of allele-specific RNAi effectors targeting genetic forms of Parkinson's disease.
Author(s): Sibley, CR
Wood, MJ
Item Type: Journal Article
Abstract: Parkinsons disease (PD) is a progressive neurological disorder affecting an estimated 5-10 million people worldwide. Recent evidence has implicated several genes that directly cause or increase susceptibility to PD. As well as advancing understanding of the genetic aetiology of PD these findings suggest new ways to modify the disease course, in some cases through genetic manipulation. Here we generated a walk-through series of RNA Pol III-expressed shRNAs targeting both the α-synuclein A30P and LRRK2 G2019S PD-associated mutations. Allele-specific discrimination of the α-synuclein A30P mutation was achieved with alignments at position 10, 13 and 14 in two model systems, including a heterozygous model mimicking the disease setting, whilst 5RACE was used to confirm stated alignments. Discrimination of the most common PD-linked LRRK2 G2019S mutation was assessed in hemizygous dual-luciferase assays and showed that alignment of the mutation opposite position 4 of the antisense species produced robust discrimination of alleles at all time points studied. Discrimination at this position was subsequently confirmed using siRNAs, where up to 10-fold discrimination was seen. The results suggest that RNAi-mediated silencing of PD-associated autosomal dominant genes could be a novel therapeutic approach for the treatment of the relevant clinical cases of PD in future.
Publication Date: 21-Oct-2011
Date of Acceptance: 22-Sep-2011
URI: http://hdl.handle.net/10044/1/30533
DOI: https://dx.doi.org/10.1371/journal.pone.0026194
ISSN: 1932-6203
Publisher: Public Library of Science
Start Page: e26194
End Page: e26194
Journal / Book Title: PLOS One
Volume: 6
Issue: 10
Copyright Statement: © 2011 Sibley, Wood. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords: Alleles
Cell Line
Cell Survival
Humans
Microscopy, Fluorescence
Mutation
Parkinson Disease
Protein-Serine-Threonine Kinases
RNA, Small Interfering
alpha-Synuclein
Cell Line
Humans
Parkinson Disease
Protein-Serine-Threonine Kinases
RNA, Small Interfering
Microscopy, Fluorescence
Cell Survival
Mutation
Alleles
alpha-Synuclein
General Science & Technology
MD Multidisciplinary
Publication Status: Published
Conference Place: United States
Appears in Collections:Department of Medicine
Faculty of Medicine



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