Double-mismatched siRNAs enhance selective gene silencing of a mutant ALS-causing allele

Acta Pharmacologica Sinica
Chang-ming Geng, Hongliu Ding

Abstract

Our previous study demonstrated an siRNA-mediated, allele-specific silencing of mutant genes that cause amyotrophic lateral sclerosis. To improve siRNA design for better therapeutic use of RNA interference, we systematically tested the base-pairing mismatch strategy in the design of asymmetric siRNA. A naturally symmetric siRNA that targets the human Cu Zn superoxide dismutase G85R mutant allele was modified by placing either 1 or 2 mismatches at the end of the siRNA from position 1 to 4 at each time. The target preference and silencing efficacy of modified siRNA were measured using a modified dual luciferase system. The modification of single base-pairing mismatch successfully achieved the conversion of the siRNA that was originally favored to the antisense of the mutant allele to the one that was favored to the sense strand of the gene. Compared to the single-mismatched siRNA, those with double-mismatch at one end demonstrated an increased asymmetry, and thus, an enhanced specificity and efficacy of gene silencing. In addition, the siRNA with double-mismatch at both ends remained in symmetry. Our results suggest the effectiveness of converting a symmetric siRNA to an asymmetric one by introducing mismatches into its structure...Continue Reading

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Citations

Dec 2, 2011·Gene Therapy·J W Lowery, V Rosen
Aug 20, 2011·PloS One·Firoz Ahmed, Gajendra P S Raghava

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