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[用于控制哺乳动物细胞功能的合成RNA技术]

[Synthetic RNA technologies to control functions of mammalian cells].

作者信息

Saito Hirohide

机构信息

Centér for IPS Cell Research and Application, Kyoto University; 53 Kawahara-cho, Shogoin, Sakyo-ku, Kioto 606-8507, Japan; The Hakubi Center for Advanced Research, Kyoto University; Yoshida-Ushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan.

出版信息

Yakugaku Zasshi. 2015;135(3):399-404. doi: 10.1248/yakushi.14-00240-4.

DOI:10.1248/yakushi.14-00240-4
PMID:25759049
Abstract

We recently succeeded in producing nanostructures made of RNA-protein (RNP) complexes. We show that RNA and the ribosomal protein L7Ae can form a triangular-like nanostructure that consists of three L7Ae proteins, which form the apices of the triangle, bound to one RNA scaffold. This shape is created through a 60° kink introduced into the RNA structure on L7Ae binding. By varying the size of the RNA scaffold we could in turn alter the overall size of the triangular nanostructure. Several functions can be added to this nanostructure by the introduction of effector proteins fused to L7Ae. The design and construction of functional RNP nanostructures that detect specific cancer cells are discussed herein. In parallel, we developed synthetic RNP translational switches to control production levels of particular proteins depending on certain input(s) within the intracellular environment. The RNP-binding module was successfully incorporated into mRNA to generate functional RNP switches. The designed ON/OFF translational switches detect expression of the trigger factor and repress or activate expression of a desired protein (e.g., apoptosis regulator) in target mammalian cells. Taken together, RNP-binding module could be employed for constructing designer genetic switches and functional nanostructures to regulate cellular processes.

摘要

我们最近成功制备了由RNA-蛋白质(RNP)复合物构成的纳米结构。我们发现RNA与核糖体蛋白L7Ae能够形成一种三角形纳米结构,该结构由三个L7Ae蛋白组成三角形的顶点,并与一个RNA支架结合。这种形状是通过在L7Ae结合时引入到RNA结构中的60°扭结形成的。通过改变RNA支架的大小,我们能够相应地改变三角形纳米结构的整体尺寸。通过引入与L7Ae融合的效应蛋白,可以为这种纳米结构添加多种功能。本文讨论了用于检测特定癌细胞的功能性RNP纳米结构的设计与构建。同时,我们开发了合成RNP翻译开关,以根据细胞内环境中的特定输入来控制特定蛋白质的产生水平。RNP结合模块已成功整合到mRNA中以生成功能性RNP开关。所设计的开/关翻译开关可检测触发因子的表达,并在目标哺乳动物细胞中抑制或激活所需蛋白质(如凋亡调节因子)的表达。综上所述,RNP结合模块可用于构建定制的基因开关和功能性纳米结构,以调节细胞过程。

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