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. 2013 Jul 24;41(18):8788–8801. doi: 10.1093/nar/gkt594

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — Mechanistic studies of abasic siRNAs. (A) HTT mRNA levels after treating with 25 nM of abasic duplex RNAs in HD patient fibroblasts (GM04281, CAG 69/17). siHdh1 is a positive control siRNA targeting HTT mRNA at a sequence outside the trinucleotide repeat region. (B) In vitro cleavage assay using RNA antisense strands and recombinant human AGO2 protein. Ladder: radiolabeled 10-nt DNA markers; NT: no treatment; REP: fully complementary anti-CAG siRNA; H153: fully complementary siRNA targeting HTT upstream region of CAG repeat; AB3 and AB6 are abasic-substituted siRNAs. (C) RNA immunoprecipitation (RIP) using an anti-AGO2 antibody reveals association of AGO2/siRNA complexes with HTT mRNA (GM04281). (D) Abasic siRNA AB8 selectively inhibits mutant HTT or ataxin-3 expression in a cooperative manner.