Rational design leads to more potent RNA interference against hepatitis B virus: factors effecting silencing efficiency.
Rational design leads to more potent RNA interference against hepatitis B virus: factors effecting silencing efficiency.
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DOI：
10.1038/mt.2008.273
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发表时间：
2009-03
影响因子：
12.4
通讯作者：
McCaffrey, Anton P.
中科院分区：
文献类型：
--
作者：
Keck, Kathy;Volper, Esther M.;Spengler, Ryan M.;Long, Dang D.;Chan, Chi Y.;Ding, Ye;McCaffrey, Anton P.
RNA干扰(RNAi)可以是一种有效的抗病毒药物；然而，RNAi的过度表达可能通过与内源性microRNA(MiRNA)机制的竞争而有毒。我们使用合理的设计来确定在无毒剂量下有效的高度有效的RNAi。对与活度相关的热力学特征进行了统计分析。选择符合与活性RNAi相关的共识内部稳定性图谱(Isp)的序列，并在内源性miRNA的背景下表达RNAi触发器。这些方法产生了高度活跃的乙肝病毒(HBVRNAi)。一项统计分析发现，通过将种子序列内的结合到目标RNA中的可访问区域，活性和成核之间存在相关性。导向链被选为有利的链偏置，但增加的链偏置与效力无关，这表明存在阈值效应。此前有报道称，外源短发夹RNA(ShRNAs)与miRNAs竞争miRNA/RNAi机制，而不是miRNAs。相反，我们证明了外源聚合酶III而不是聚合酶II驱动的miRNAs在miRNA途径的多个步骤中与外源miRNAs竞争。外源miRNAs也与内源miR-21竞争。因此，即使在使用基于miRNA的疗法时，也应该监测与内源性miRNA的竞争。然而，在没有观察到竞争的剂量下，实现了有效的沉默。
RNA interference (RNAi) can be an effective antiviral agent; however, overexpression of RNAi can be toxic through competition with the endogenous microRNA (miRNA) machinery. We used rational design to identify highly potent RNAi that is effective at nontoxic doses. A statistical analysis was conducted to pinpoint thermodynamic characteristics correlated with activity. Sequences were selected that conformed to a consensus internal stability profile (ISP) associated with active RNAi, and RNAi triggers were expressed in the context of an endogenous miRNA. These approaches yielded highly active hepatitis B virus (HBV) RNAi. A statistical analysis found a correlation between activity and nucleation by binding within the seed sequence to accessible regions in the target RNA. Guide strands were selected for favorable strand biasing, but increased strand biasing did not correlate with potency, suggesting a threshold effect. Exogenous short hairpin RNAs (shRNAs), but not miRNAs were previously reported to compete with miRNAs for the miRNA/RNAi machinery. In contrast, we show that exogenous Polymerase III- but not Polymerase II-driven miRNAs compete with exogenous miRNAs, at multiple steps in the miRNA pathway. Exogenous miRNAs also compete with endogenous miR-21. Thus, competition with endogenous miRNAs should be monitored even when using miRNA-based therapeutics. However, potent silencing was achieved at doses where competition was not observed.
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影响因子：
14.9
作者：
Ding, Y;Chan, CY;Lawrence, CE
通讯作者：
Lawrence, CE
影响因子：
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作者：
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