3′ UTR seed matches, but not overall identity, are associated with RNAi off-targets
3′ UTR seed matches, but not overall identity, are associated with RNAi off-targets
复制标题
DOI：
10.1038/nmeth854
复制
发表时间：
2006-03-01
期刊：
影响因子：
48
通讯作者：
Khvorova, A
中科院分区：
文献类型：
--
作者：
Birmingham, A;Anderson, EM;Khvorova, A
脱靶基因沉默在解释来自大规模RNA干扰（RNAi）筛选的数据方面可能存在显著挑战。我们对通过小干扰RNA（siRNA）转染的人细胞表达谱鉴定的脱靶基因进行了详细分析。与通常的假设相反，对随后的脱靶基因数据库的分析表明，除了接近完美的匹配之外，总体同一性对确定特定基因的表达是否会受到给定siRNA的影响几乎没有贡献。相反，脱靶与存在一个或多个与siRNA的反义链的六聚体或七聚体种子区（位置2-7或2-8）完全匹配的3'非翻译区（UTR）有关。这些发现对未来的siRNA设计以及RNAi在高通量筛选和治疗开发中的应用具有重要意义。
Off-target gene silencing can present a notable challenge in the interpretation of data from large-scale RNA interference (RNAi) screens. We performed a detailed analysis of off-targeted genes identified by expression profiling of human cells transfected with small interfering RNA (siRNA). Contrary to common assumption, analysis of the subsequent off-target gene database showed that overall identity makes little or no contribution to determining whether the expression of a particular gene will be affected by a given siRNA, except for near-perfect matches. Instead, off-targeting is associated with the presence of one or more perfect 3' untranslated region (UTR) matches with the hexamer or heptamer seed region (positions 2-7 or 2-8) of the antisense strand of the siRNA. These findings have strong implications for future siRNA design and the application of RNAi in high-throughput screening and therapeutic development.