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A study on the fundamental factors determining the efficacy of siRNAs with high C/G contents

机译：影响高C / G含量的siRNA效能的基本因素研究

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摘要

Although there are many reports about the efficacy of siRNAs, it is not clear whether those siRNAs with high C/G contents can be used to silence their target mRNAs efficiently. In this study, we investigated the structure and function of a group of siRNAs with high C/G contents. The results showed that single siRNAs against the Calpain, Otoferlin and Her2 mRNAs could induce different silencing effects on their targets, suggesting that the accessibility to target sequences influences the efficacy of siRNA. Unexpectedly, a single siRNA could target its cognate sequence in the 3’UTR of EEF1D or the 5’UTR of hTRF2 or CDC6. Their interaction induced different modes of gene silencing. Furthermore, the introduction of mutations into the 3’ end of the passenger strand showed that the position and number of mutated nucleotides could exert some influence on the efficacy of siRNA. However, these mutations did not completely block the passenger strand from exerting its RNAi effect. Interestingly, our findings also indicated that the target mRNA might play essential roles in maintaining or discarding the guide strand in RISCs. Thus, the conclusion could be drawn that favorable siRNA sequences, accessible target structures and the fast cleavage mode are necessary and sufficient prerequisites for efficient RNAi.
机译：尽管有许多关于siRNA功效的报道，但尚不清楚这些C / G含量高的siRNA是否可用于有效地沉默其靶mRNA。在这项研究中，我们调查了一组具有高C / G含量的siRNA的结构和功能。结果表明，针对Calpain，Otoferlin和Her2 mRNA的单个siRNA可能对其靶标产生不同的沉默作用，这表明靶标序列的可及性影响siRNA的功效。出乎意料的是，单个siRNA可能在EEF1D的3'UTR或hTRF2或CDC6的5'UTR中靶向其同源序列。它们的相互作用诱导了基因沉默的不同模式。此外，在过客链3'端引入突变表明，突变核苷酸的位置和数量可能对siRNA的功效产生一些影响。但是，这些突变并没有完全阻止过客链发挥其RNAi效应。有趣的是，我们的发现还表明，靶标mRNA可能在维持或丢弃RISC中的引导链中起重要作用。因此，可以得出结论，有利的siRNA序列，可及的靶结构和快速切割模式是有效RNAi的必要和充分前提。

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