...
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Thermodynamic stability of small hairpin RNAs highly influences the loading process of different mammalian Argonautes
【24h】

Thermodynamic stability of small hairpin RNAs highly influences the loading process of different mammalian Argonautes

机译:小发夹RNA的热力学稳定性极大地影响了不同哺乳动物Argonautes的加载过程

获取原文
获取原文并翻译 | 示例
           

摘要

MicroRNAs and siRNAs interact with target sequences in mRNAs, inducing cleavage- and non-cleavage-based gene repression through the RNA-induced silencing complex (RISC) that consists of one of four mammalian Argonaute proteins, Ago1-Ago4. The process of how Dicer substrate small hairpin RNAs (shRNAs) are loaded into different mammalian Agos in vivo is not well established. Here we report that shRNAs are loaded into mammalian Agos in two stepwise processes, physical association and activation, with the latter being the rate-limiting step with noncleaving RISC. We establish that, although RNA duplexes processed from shRNAs bind to Agos in cells with similar affinity, the degree by which the complexes are activated (coupled with the removal of the passenger strand) correlates with the thermodynamic instability of RNA duplexes being loaded rather than the structure of the RNA, as was previously demonstrated in Drosophila. Interestingly, Ago loading of siRNAs is less sensitive to thermostability than that of their shRNA equivalents. These results may have important implications for the future design of RNAi-based therapeutics.
机译:MicroRNA和siRNA与mRNA中的靶序列相互作用,通过RNA诱导的沉默复合物(RISC)诱导基于切割和非切割的基因阻抑,该复合物由四个哺乳动物Argonaute蛋白Ago1-Ago4之一组成。 Dicer底物小发夹RNA(shRNA)如何在体内加载到不同的哺乳动物Agos的过程尚不完善。在这里,我们报告说,shRNA通过两个逐步的过程(物理关联和激活)被加载到哺乳动物Agos中,后者是非裂解RISC的限速步骤。我们确定,尽管从shRNA加工的RNA双链体以相似的亲和力与细胞中的Agos结合,但复合体被激活的程度(与去除客链的结合)与被加载的RNA双链体的热力学不稳定性相关,而不是与如先前在果蝇中所证实的,RNA的结构。有趣的是,siRNA的Ago加载对热稳定性的敏感性低于其shRNA等同物。这些结果可能对基于RNAi的疗法的未来设计具有重要意义。

著录项

  • 来源
  • 作者单位

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

    Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010;

    Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    rna interference; gene silencing;

    机译:RNA干扰;基因沉默;

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号