Chemical Modification of the siRNA Seed Region Suppresses Off-Target Effects by Steric Hindrance to Base-Pairing with Targets.
Chemical Modification of the siRNA Seed Region Suppresses Off-Target Effects by Steric Hindrance to Base-Pairing with Targets.
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DOI：
10.1021/acsomega.7b00291
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发表时间：
2017-05-31
期刊：
影响因子：
4.1
通讯作者：
Ui-Tei K
中科院分区：
文献类型：
--
作者：
Iribe H;Miyamoto K;Takahashi T;Kobayashi Y;Leo J;Aida M;Ui-Tei K
小干扰RNA（siRNA）引导链的种子区（位置2-8）中的核苷酸的2′-O-甲基（2′-OMe）和锁核酸（LNA）的化学修饰显著降低了种子匹配（SM）脱靶效应。具有2′-OMe修饰的siRNA抑制完全匹配（CM）靶基因的表达，而具有LNA修饰的siRNA不抑制CM靶基因的表达。通过这些修饰对siRNA构象变化的计算预测，我们发现siRNA种子区的两种修饰都通过与靶转录物碱基配对的空间位阻减少了SM脱靶效应，但LNA修饰也通过改变RNA构象干扰了siRNA引导链与Argonaute（AGO）蛋白的结合。因此，siRNA引导链的化学修饰（其改变空间构象以干扰与靶转录物的碱基配对，但不干扰与AGO蛋白的缔合）可以成功地抑制脱靶效应，而不显著损失RNA沉默活性。
Chemical modifications of 2′-O-methyl (2′-OMe) and locked nucleic acid (LNA) of the nucleotides in the seed region (positions 2–8) of the small interfering RNA (siRNA) guide strand significantly reduced seed-matched (SM) off-target effects. The siRNA with 2′-OMe modifications inhibited the expression of a completely-matched (CM) target gene, whereas that with LNA modifications did not inhibit the expression of the CM target. By computational predictions of conformational changes of siRNA by these modifications, we revealed that both modifications in the siRNA seed region reduce SM off-target effects by steric hindrance to base-pairing with target transcripts but LNA modifications also disturb the association of the siRNA guide strand with the Argonaute (AGO) protein by altering RNA conformation. Thus, chemical modifications of the siRNA guide strand, which alter steric conformation to disturb base-pairing with target transcripts but do not disturb the association with the AGO protein, may successfully suppress off-target effects without substantial loss of RNA silencing activity.
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