Endless: a purine-binding RNA motif that can be expressed in cells.

Endless: a purine-binding RNA motif that can be expressed in cells.
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Endless:可在细胞中表达的嘌呤结合 RNA 基序

DOI:
10.1002/anie.201403579
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发表时间:
2014
期刊:
影响因子:
--
通讯作者:
C. Richert
C. Richert
中科院分区:
--
文献类型:
--
作者:
C. Kröner;M. Thunemann;S. Vollmer;M. Kinzer;R. Feil;C. Richert

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越来越清楚的是,自然界广泛使用RNA来调节细胞的重要功能,并且短序列经常用于抑制基因表达。然而,通过设计折叠成配体结合结构的RNA序列来控制细胞内小分子的浓度是困难的。描述了“无尽”的开发,这是一种基于三链体的折叠基序,可在哺乳动物细胞中表达并结合第二信使3′,5 ′-环鸟苷一磷酸(cGMP)。在体外,DNA或RNA版本的endless显示出低的微摩尔至纳摩尔的cGMP解离常数。 为了测试其在体内的功能性,在小鼠血管平滑肌细胞中,将串联排列的无四链RNA基序与荧光cGMP传感器蛋白共表达。 与表达对照基序的细胞相比,一氧化氮诱导的内源性cGMP信号在无限表达细胞中受到抑制,这表明无限表达可以作为基因编码的cGMP接收器来调节细胞中的信号转导。
It is becoming increasingly clear that nature uses RNAs extensively for regulating vital functions of the cell, and short sequences are frequently used to suppress gene expression. However, controlling the concentration of small molecules intracellularly through designed RNA sequences that fold into ligand‐binding structures is difficult. The development of “endless”, a triplex‐based folding motif that can be expressed in mammalian cells and binds the second messenger 3′,5′‐cyclic guanosine monophosphate (cGMP), is described. In vitro, DNA or RNA versions ofendlessshow low micromolar to nanomolar dissociation constants for cGMP. To test its functionality in vivo, fourendlessRNA motifs arranged in tandem were co‐expressed with a fluorescent cGMP sensor protein in murine vascular smooth muscle cells. Nitric oxide induced endogenous cGMP signals were suppressed inendless‐expressing cells compared to cells expressing a control motif, which suggests thatendlesscan act as a genetically encoded cGMP sink to modulate signal transduction in cells.
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