Relative Quantification of siRNA Strand Loading into Ago2 for Design of Highly Active siRNAs
Relative Quantification of siRNA Strand Loading into Ago2 for Design of Highly Active siRNAs
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DOI：
10.1007/978-1-4939-9220-1_4
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发表时间：
2019-01-01
期刊：
影响因子：
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通讯作者：
Walton, S. Patrick
中科院分区：
文献类型：
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作者：
Angart, Phillip A.;Adu-Berchie, Kwasi;Walton, S. Patrick
在RNA干扰（RNAi）中，沉默是通过双链小干扰RNA（siRNA）与必要的RNAi途径蛋白（包括Argonaute 2（Ago2））相互作用来实现的。基于这些相互作用，siRNA的一条链被加载到Ago2中，形成活性RNA诱导的沉默复合物（RISC）。最佳siRNA使针对预期靶标的RISC活性最大化，并使脱靶沉默最小化。为了实现所需的活性和特异性，选择合适的siRNA链加载到Ago 2中至关重要。在这里，我们提供了一个方案来量化单个siRNA链到Ago2中的相对负载，这是确定siRNA实现沉默活性和靶特异性的能力的一个因素。
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.