siRNA off-target effects can be reduced at concentrations that match their individual potency.
siRNA off-target effects can be reduced at concentrations that match their individual potency.
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DOI：
10.1371/journal.pone.0021503
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Hughes JD
中科院分区：
文献类型：
--
作者：
Caffrey DR;Zhao J;Song Z;Schaffer ME;Haney SA;Subramanian RR;Seymour AB;Hughes JD
小干扰rna （sirna）通常用于降低特定基因的mRNA水平，目的是研究其功能。几项研究表明，sirna并不总是特异性的，而且可能有许多脱靶效应。脱靶mrna的3 ' utr通常富集在与siRNA种子区互补的序列中。我们证明，当细胞用相对较低的siRNA剂量（例如1 nM）处理时，siRNA脱靶可以显着减少，但足以有效地沉默预期的靶标。对于靶向STAT3或己糖激酶II的修饰和未修饰的sirna，脱靶量的减少都得到了证明。低浓度降低了与siRNA种子区互补的转录本的沉默。同样，不与siRNA互补的脱靶基因在较低剂量下减少，包括参与免疫反应的上调基因。重要的是，用靶向己糖激酶II的siRNA治疗后，意外诱导caspase活性也被证明是一种浓度依赖性脱靶效应。我们的结论是，某些siRNA可以在低浓度下沉默其预期靶标，从而减少脱靶及其相关的表型效应。
Small interfering RNAs (siRNAs) are routinely used to reduce mRNA levels for a specific gene with the goal of studying its function. Several studies have demonstrated that siRNAs are not always specific and can have many off-target effects. The 3′ UTRs of off-target mRNAs are often enriched in sequences that are complementary to the seed-region of the siRNA. We demonstrate that siRNA off-targets can be significantly reduced when cells are treated with a dose of siRNA that is relatively low (e.g. 1 nM), but sufficient to effectively silence the intended target. The reduction in off-targets was demonstrated for both modified and unmodified siRNAs that targeted either STAT3 or hexokinase II. Low concentrations reduced silencing of transcripts with complementarity to the seed region of the siRNA. Similarly, off-targets that were not complementary to the siRNA were reduced at lower doses, including up-regulated genes that are involved in immune response. Importantly, the unintended induction of caspase activity following treatment with a siRNA that targeted hexokinase II was also shown to be a concentration-dependent off-target effect. We conclude that off-targets and their related phenotypic effects can be reduced for certain siRNA that potently silence their intended target at low concentrations.
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影响因子：
46.9
作者：
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通讯作者：
Linsley, PS
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作者：
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通讯作者：
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