Naturally occurring three-way junctions can be repurposed as genetically encoded RNA-based sensors.
Naturally occurring three-way junctions can be repurposed as genetically encoded RNA-based sensors.
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DOI：
10.1016/j.chembiol.2021.04.022
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发表时间：
2021-11-18
影响因子：
8.6
通讯作者：
Jaffrey SR
中科院分区：
文献类型：
--
作者：
Moon JD;Wu J;Dey SK;Litke JL;Li X;Kim H;Jaffrey SR
使用由RNA组成的生物传感器可以在活细胞中对小分子进行成像。然而，基于RNA的设备很难设计。在这里，我们描述了一个通用的平台，用于设计基于RNA的荧光小分子传感器，使用天然存在的高度稳定的三向连接RNA。我们表明，配体结合适体和荧光适体可以插入到三路交界处，并连接的方式，使三路交界处作为一个小分子调控的荧光传感器在体外和细胞。传感器被设计成使得三向接合中的螺旋间稳定相互作用仅在配体结合时被诱导。我们使用这些基于RNA的设备来测量哺乳动物细胞中S-腺苷甲硫氨酸水平的动态，真实的时间。我们表明，这种策略是兼容不同的代谢物结合RNA适体，荧光适体，和三通路口。总的来说，这些数据证明了一种用于容易地产生在活细胞中起作用的RNA装置的通用方法。基因编码RNA生物传感器是一种新兴的工具，用于成像细胞中的小分子代谢物。在这份手稿中，Moon et.等人描述了一种用于开发RNA生物传感器的通用方法，该方法通过经再工程化的天然存在的RNA三向接头连接荧光RNA适体和代谢物结合适体。
Small molecules can be imaged in living cells using biosensors composed of RNA. However, RNA-based devices are difficult to design. Here we describe a versatile platform for designing RNA-based fluorescent small molecule sensors using naturally occurring highly stable three-way junction RNAs. We show that ligand-binding aptamers and fluorogenic aptamers can be inserted into three-way junctions, and connected in a way that enables the three-way junction to function as a small molecule-regulated fluorescent sensor in vitro and in cells. The sensors are designed so that the inter-helical stabilizing interactions in the three-way junction are only induced upon ligand binding. We use these RNA-based devices to measure the dynamics of S-adenosylmethionine levels in mammalian cells in real time. We show that this strategy is compatible with diverse metabolite-binding RNA aptamers, fluorogenic aptamers, and three-way junctions. Overall, these data demonstrate a versatile method for readily generating RNA-devices that function in living cells. Genetically encoded RNA biosensors are an emerging tool for imaging small-molecule metabolites in cells. In this manuscript, Moon et. al. describe a versatile method for developing RNA biosensors by connecting fluorogenic RNA aptamers and metabolite-binding aptamers via reengineered, naturally occurring RNA three-way junctions.
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影响因子：
3.2
作者：
ALATOSSAVA, T;JUTTE, H;KELLENBERGER, E
通讯作者：
KELLENBERGER, E
影响因子：
4.5
作者：
Lescoute, A;Westhof, E
通讯作者：
Westhof, E
影响因子：
8.6
作者：
Kim, Hyaeyeong;Jaffrey, Samie R.
通讯作者：
Jaffrey, Samie R.
DOI：
10.1073/pnas.1016690108
发表时间：
2011-05-03
影响因子：
11.1
作者：
Ding, Fang;Lu, Changrui;Ke, Ailong
通讯作者：
Ke, Ailong
含有 G-四链体的 RNA 会激活类 GFP 荧光团中的荧光。
影响因子：
14.8
作者：
Huang, Hao;Suslov, Nikolai B.;Li, Nan-Sheng;Shelke, Sandip A.;Evans, Molly E.;Koldobskaya, Yelena;Rice, Phoebe A.;Piccirilli, Joseph A.
通讯作者：
Piccirilli, Joseph A.