Dynamic signal processing by ribozyme-mediated RNA circuits to control gene expression

Dynamic signal processing by ribozyme-mediated RNA circuits to control gene expression
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通过核酶介导的 RNA 电路进行动态信号处理来控制基因表达

DOI:
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发表时间:
2015
期刊:
bioRxiv
影响因子:
--
通讯作者:
A. Jaramillo
A. Jaramillo
中科院分区:
--
文献类型:
--
作者:
Shensi Shen;G. Rodrigo;S. Prakash;E. Majer;Thomas E. Landrain;B. Kirov;J. Daròs;A. Jaramillo

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生物体有不同的电路,可以将信号分子水平转化为基因表达的变化。合成生物学中的一个重要挑战涉及RNA模块的重新设计,使活细胞中的动态信号处理成为可能。这需要一种可扩展的传感、传输和驱动方法,可以组装成更大的信号网络。在这里,我们提出了一种生化策略来设计rna介导的信号转导级联,能够感知小分子和小rna。我们利用链置换技术设计了可切换的功能性RNA结构域。我们通过实验表征了合成RNA信号级联的分子机制,展示了通过转导RNA信号调节基因表达的能力,并描述了我们的系统对单个活细胞周期性强迫的信号处理响应。该工程系统将RNA-RNA相互作用与可用的核酶和适体元件相结合,为设计任意复杂的基因回路提供了新方法。
Organisms have different circuitries that allow converting signal molecule levels to changes in gene expression. An important challenge in synthetic biology involves the de novo design of RNA modules enabling dynamic signal processing in live cells. This requires a scalable methodology for sensing, transmission, and actuation, which could be assembled into larger signaling networks. Here, we present a biochemical strategy to design RNA-mediated signal transduction cascades able to sense small molecules and small RNAs. We design switchable functional RNA domains by using strand-displacement techniques. We experimentally characterize the molecular mechanism underlying our synthetic RNA signaling cascades, show the ability to regulate gene expression with transduced RNA signals, and describe the signal processing response of our systems to periodic forcing in single live cells. The engineered systems integrate RNA-RNA interaction with available ribozyme and aptamer elements, providing new ways to engineer arbitrary complex gene circuits.
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