Tolerance for mutations and chemical modifications in a siRNA
Tolerance for mutations and chemical modifications in a siRNA
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DOI：
10.1093/nar/gkg147
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发表时间：
2003-01-15
影响因子：
14.9
通讯作者：
Prydz, H
中科院分区：
文献类型：
--
作者：
Amarzguioui, M;Holen, T;Prydz, H
短干扰RNA（Short interfering RNA，siRNA）是RNA干扰的活性物质，在基础和临床研究中具有重要的应用前景。我们探索了靶向凝血起始剂组织因子的siRNA的两条21-nt链的各个部分对突变和化学修饰的耐受性。突变为G/C颠换。化学修饰为2 ′-O-甲基化、2 ′-O-烯丙基化和硫代磷酸酯化。我们发现siRNA通常耐受5'端的突变，而3'端表现出低耐受性。这一观察结果可能有助于设计特异性靶向含有单核苷酸多态性的转录本的siRNA。我们进一步证明，在我们的系统中，野生型siRNA的单个反义链几乎与siRNA双链体一样有效，而相应的甲基化M2+4形式的反义具有降低的活性。测试的大多数化学修饰版本具有接近野生型的初始活性，而某些siRNA种类的长期活性增加。我们的研究结果可能会改善设计的siRNA在体内实验。
Short interfering RNA (siRNA), the active agent of RNA interference, shows promise of becoming a valuable tool in both basic and clinical research. We explore the tolerance to mutations and chemical modifications in various parts of the two 21-nt strands of a siRNA targeting the blood clotting initiator Tissue Factor. The mutations were G/C transversions. The chemical modifications were 2'-O-methylation, 2'-O-allylation and phosphorothioates. We found that siRNA generally tolerated mutations in the 5' end, while the 3' end exhibited low tolerance. This observation may facilitate the design of siRNA for specific targeting of transcripts containing single nucleotide polymorphisms. We further demonstrate that in our system the single antisense strand of the wild-type siRNA is almost as effective as the siRNA duplex, while the corresponding methylated M2+4 version of the antisense had reduced activity. Most of the chemically modified versions tested had near-wild-type initial activity, while the long-term activity was increased for certain siRNA species. Our results may improve the design of siRNAs for in vivo experiments.