Abstract
In RNA interference (RNAi), silencing is achieved through the interaction of double-stranded small interfering RNAs (siRNAs) with essential RNAi pathway proteins, including Argonaute 2 (Ago2). Based on these interactions, one strand of the siRNA is loaded into Ago2 forming the active RNA-induced silencing complex (RISC). Optimal siRNAs maximize RISC activity against the intended target and minimize off-target silencing. To achieve the desired activity and specificity, selection of the appropriate siRNA strand for loading into Ago2 is essential. Here, we provide a protocol to quantify the relative loading of individual siRNA strands into Ago2, one factor in determining the capacity of a siRNA to achieve silencing activity and target specificity.
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Angart, P.A., Adu-Berchie, K., Carlson, R.J., Vocelle, D.B., Chan, C., Walton, S.P. (2019). Relative Quantification of siRNA Strand Loading into Ago2 for Design of Highly Active siRNAs. In: Dinesh Kumar, L. (eds) RNA Interference and Cancer Therapy. Methods in Molecular Biology, vol 1974. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9220-1_4
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DOI: https://doi.org/10.1007/978-1-4939-9220-1_4
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