Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy
Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy
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DOI：
10.1038/nbt1051
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发表时间：
2005-02-01
影响因子：
46.9
通讯作者：
Rossi, JJ
中科院分区：
文献类型：
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作者：
Kim, DH;Behlke, MA;Rossi, JJ
RNA干扰（RNAi）是由双链RNA 1 -3引发的序列特异性转录后基因沉默的过程。在试图鉴定在较低浓度下有效发挥作用的RNAi触发剂时，我们发现长度为25-30个核苷酸的合成RNA双链体的效力比相应的常规21-mer小干扰RNA（siRNA）高100倍。一些21-mer siRNA难以沉默的位点可以被27-mer双链体有效靶向，沉默持续长达10 d。值得注意的是，27-mer不诱导干扰素或激活蛋白激酶R（PKR）。较长的双链体的增强的效力归因于它们是Dicer内切核酸酶的底物的事实，直接将siRNA的产生与RNA诱导的沉默复合物中的掺入联系起来。这些结果提供了使用低浓度的合成RNA作为细胞Dicer介导的切割的底物来引发RNAi介导的靶切割的替代策略。
RNA interference (RNAi) is the process of sequence-specific post-transcriptional gene silencing triggered by double-stranded RNAs1-3. In attempts to identify RNAi triggers that effectively function at lower concentrations, we found that synthetic RNA duplexes 25-30 nucleotides in length can be up to 100-fold more potent than corresponding conventional 21-mer small interfering RNAs (siRNAs). Some sites that are refractory to silencing by 21-mer siRNAs can be effectively targeted by 27-mer duplexes, with silencing lasting up to 10 d. Notably, the 27-mers do not induce interferon or activate protein kinase R (PKR). The enhanced potency of the longer duplexes is attributed to the fact that they are substrates of the Dicer endonuclease, directly linking the production of siRNAs to incorporation in the RNA-induced silencing complex. These results provide an alternative strategy for eliciting RNAi-mediated target cleavage using low concentrations of synthetic RNA as substrates for cellular Dicer-mediated cleavage.