TY  - JOUR
IS  - 10
N1  - © 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.

Funding: This work was supported a Medical Research Council Studentship (CRS). This work was supported by a research grants from Parkinson's UK to MJAW. A free gift of an siRNA was provided by Novartis Pharmaceuticals with nothing in exchange. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
AV  - public
VL  - 6
Y1  - 2011/10/21/
TI  - Identification of allele-specific RNAi effectors targeting genetic forms of Parkinson's disease.
A1  - Sibley, CR
A1  - Wood, MJ
KW  - Alleles
KW  -  Cell Line
KW  -  Cell Survival
KW  -  Humans
KW  -  Microscopy
KW  -  Fluorescence
KW  -  Mutation
KW  -  Parkinson Disease
KW  -  Protein-Serine-Threonine Kinases
KW  -  RNA
KW  -  Small Interfering
KW  -  alpha-Synuclein
JF  - PLoS One
UR  - http://dx.doi.org/10.1371/journal.pone.0026194
SN  - 1932-6203
N2  - Parkinson's 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 ?-synuclein A30P and LRRK2 G2019S PD-associated mutations. Allele-specific discrimination of the ?-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 5'RACE 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.
ID  - discovery1392155
ER  -