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Scott R Suter¹, Jessica Sheu-Gruttadauria², Nicole T Schirle²

¹Department of Chemistry, University of California, Davis , One Shields Avenue, Davis, California 95616, United States.

Journal of the American Chemical Society

|July 9, 2016

概括

化学修饰的短干扰RNAs (siRNAs) 显示出改善的治疗潜力. siRNA中的新型三基核酸增强了向特异性并减少了非向效应,为药物开发提供了新的策略.

科学领域:

生物化学
分子生物学
药物开发

背景情况:

短干扰RNAs (siRNAs) 是利用RNA干扰 (RNAi) 途径的有价值的治疗方法.
化学修饰对于克服原生siRNAs在药物开发中的结构限制至关重要.
对人类RNAi路径的结构洞察力有助于优化siRNA特性.

研究的目的:

确定与修饰的siRNA结合的人类阿尔戈诺特2 (hAgo2) 的结构.
研究1 (g1) 位置的三基核酸如何影响siRNA的结合和功能.
探索结构引导的修改来提高siRNA的选择性和效力.

主要方法:

用X射线结晶学确定Hago2的2.3 Å分辨率结构与g1修饰的siRNA.
生物化学测试以测量修饰的siRNA与向RNA的结合亲和力.
功能性测试,以评估目标和目标之外的抗击效果.

主要成果:

三基核酸模拟物插入到 hAgo2 中央RNA结合裂中,调节导向-目标RNA配对.
修改显著降低了对只有种子匹配的目标的结合亲和力,但并没有完全匹配的目标.
减少了非位抑制的siRNA功效 (IC50增加了4倍),同时改善了位抑制 (IC50减少了2倍).

结论:

在siRNA中引入非自然的核酸,如三类型,提供一种基于结构的方法来控制标的结合和强度.
这种修改通过减少目标外效应,同时改善目标基因沉默来提高siRNA的选择性.
将替代组投射到 hAgo2 中心裂为开发更有选择性的siRNA疗法提供了一个新的策略.

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