Genetically Encoded Catalytic Hairpin Assembly for Sensitive RNA Imaging in Live Cells.
Genetically Encoded Catalytic Hairpin Assembly for Sensitive RNA Imaging in Live Cells.
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DOI：
10.1021/jacs.8b03956
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发表时间：
2018-07-18
影响因子：
15
通讯作者：
You M
中科院分区：
文献类型：
--
作者：
Karunanayake Mudiyanselage APKK;Yu Q;Leon-Duque MA;Zhao B;Wu R;You M
DNA和RNA纳米技术已被用于动态分子器件的开发。特别是，可编程的无酶核酸电路，如催化发夹组装，已被证明是有用的工具，用于生物分析，并扩大系统的复杂性，在一定程度上超越目前的细胞遗传电路。然而，大多数合成核酸回路的细胞内功能受到生物递送和降解方面的挑战的阻碍。另一方面，基因编码和转录的RNA电路成为长期嵌入细胞分析和调控的替代强大工具。在此，我们报道了一种用于活细胞内敏感RNA成像的基于基因编码RNA的催化发夹组装电路。使用荧光RNA适体花椰菜的分裂版本作为报告基因。一个靶RNA可以催化触发数十到数百个西兰花的荧光。结果，可以灵敏地检测靶RNA。我们进一步设计了我们的电路，以允许轻松编程来成像各种靶RNA序列。这一设计原理为开发用于细胞应用的多种遗传编码RNA电路打开了竞技场。
DNA and RNA nanotechnology has been used for the development of dynamic molecular devices. In particular, programmable enzyme-free nucleic acid circuits, such as catalytic hairpin assembly, have been demonstrated as useful tools for bioanalysis and to scale up system complexity to an extent beyond current cellular genetic circuits. However, the intracellular functions of most synthetic nucleic acid circuits have been hindered by challenges in the biological delivery and degradation. On the other hand, genetically encoded and transcribed RNA circuits emerge as alternative powerful tools for long-term embedded cellular analysis and regulation. Herein, we reported a genetically encoded RNA-based catalytic hairpin assembly circuit for sensitive RNA imaging inside living cells. The split version of Broccoli, a fluorogenic RNA aptamer, was used as the reporter. One target RNA can catalytically trigger the fluorescence from tens-to-hundreds of Broccoli. As a result, target RNAs can be sensitively detected. We have further engineered our circuit to allow easy programming to image various target RNA sequences. This design principle opens the arena for developing a large variety of genetically encoded RNA circuits for cellular applications.
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无酶DNA电路对多种检测方法的有理模块化适应。
影响因子：
14.9
作者：
Li B;Ellington AD;Chen X
通讯作者：
Chen X
DOI：
10.1126/science.1207339
发表时间：
2011-07-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Paige JS;Wu KY;Jaffrey SR
通讯作者：
Jaffrey SR
影响因子：
14.9
作者：
Srinivas N;Ouldridge TE;Sulc P;Schaeffer JM;Yurke B;Louis AA;Doye JP;Winfree E
通讯作者：
Winfree E
影响因子：
6.4
作者：
Negrete A;Shiloach J
通讯作者：
Shiloach J
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.