Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference
Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference
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哺乳动物和小鸡 RNA 干扰高效 siRNA 序列的选择指南
DOI：
10.1093/nar/gkh247
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发表时间：
2004-02-01
影响因子：
14.9
通讯作者：
Saigo, K
中科院分区：
文献类型：
--
作者：
Ui-Tei, K;Naito, Y;Saigo, K
在本研究中，短干扰RNA（siRNA）序列和RNA干扰（RNAi）效应之间的关系进行了广泛的分析，使用62个目标的4个外源基因和2个内源基因和3个哺乳动物和果蝇细胞。我们提出的规则，可以支配siRNA序列的偏好，并根据系统的哺乳动物功能基因组学所必需的高效siRNA可以很容易地设计。这些规则表明，同时满足以下所有四个序列条件的siRNA能够在哺乳动物细胞中诱导高度有效的基因沉默：（i）反义链5 ′末端的A/U;（ii）有义链5 ′末端的G/C;（iii）反义链5 ′末端三分之一处的至少五个A/U残基;和（iv）不存在任何长度超过9 nt的GC延伸。与前三种条件特征相反的SiRNA在哺乳动物细胞中几乎不引起基因沉默。发现siRNA序列偏好的基本上相同的规则适用于哺乳动物细胞中基于DNA的RNAi和使用鸡胚的卵内RNAi。与哺乳动物和鸡细胞相比，在果蝇体内RNAi中可以检测到很少的siRNA序列偏好。
In the present study, the relationship between short interfering RNA (siRNA) sequence and RNA interference (RNAi) effect was extensively analyzed using 62 targets of four exogenous and two endogenous genes and three mammalian and Drosophila cells. We present the rules that may govern siRNA sequence preference and in accordance with which highly effective siRNAs essential for systematic mammalian functional genomics can be readily designed. These rules indicate that siRNAs which simultaneously satisfy all four of the following sequence conditions are capable of inducing highly effective gene silencing in mammalian cells: (i) A/U at the 5' end of the antisense strand; (ii) G/C at the 5' end of the sense strand; (iii) at least five A/U residues in the 5' terminal one-third of the antisense strand; and (iv) the absence of any GC stretch of more than 9 nt in length. siRNAs opposite in features with respect to the first three conditions give rise to little or no gene silencing in mammalian cells. Essentially the same rules for siRNA sequence preference were found applicable to DNA-based RNAi in mammalian cells and in ovo RNAi using chick embryos. In contrast to mammalian and chick cells, little siRNA sequence preference could be detected in Drosophila in vivo RNAi.