Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules.
Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules.
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DOI：
10.1093/nar/gku655
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发表时间：
2014-08
影响因子：
14.9
通讯作者：
Yung LY
中科院分区：
文献类型：
--
作者：
Ang YS;Yung LY
生物分子相互作用具有重要的细胞意义，然而，当前的分子工具箱中缺乏一种用于感测此类邻近事件的简单方法。我们设计了一种动态 DNA 电路，能够识别邻近的目标，启动预编程的信号转导过程，从而导致局部信号放大。整个电路被设计为独立的，即它可以自主地自组装到单个目标分子上，并以最小的串扰形成局部信号。 α-凝血酶用作模型蛋白，以评估各个模块的性能以及在生理相关缓冲条件下邻近相互作用的整体电路。该电路在存在非特异性蛋白质和干扰血清基质的情况下实现了良好的选择性，并在单个混合步骤中成功检测到生理相关的 α-凝血酶浓度 (50 nM–5 μM)，无需进一步清洗。通过控制温度和探针浓度，可以在动力学上增强相互作用位点局部信号的形成。这项工作提供了一个基本的通用框架，其他电路模块可以根据该框架进行调整，用于感测其他感兴趣的生物分子或细胞相互作用。
Biomolecular interactions have important cellular implications, however, a simple method for the sensing of such proximal events is lacking in the current molecular toolbox. We designed a dynamic DNA circuit capable of recognizing targets in close proximity to initiate a pre-programmed signal transduction process resulting in localized signal amplification. The entire circuit was engineered to be self-contained, i.e. it can self-assemble onto individual target molecules autonomously and form localized signal with minimal cross-talk. α-thrombin was used as a model protein to evaluate the performance of the individual modules and the overall circuit for proximity interaction under physiologically relevant buffer condition. The circuit achieved good selectivity in presence of non-specific protein and interfering serum matrix and successfully detected for physiologically relevant α-thrombin concentration (50 nM–5 μM) in a single mixing step without any further washing. The formation of localized signal at the interaction site can be enhanced kinetically through the control of temperature and probe concentration. This work provides a basic general framework from which other circuit modules can be adapted for the sensing of other biomolecular or cellular interaction of interest.
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影响因子：
16.6
作者：
通讯作者：
--
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
影响因子：
38.3
作者：
通讯作者：
--
DOI：
10.1186/bcr2131
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Deng G;Herrler M;Burgess D;Manna E;Krag D;Burke JF
通讯作者：
Burke JF
DOI：
10.1073/pnas.0407024101
发表时间：
2004-10-26
影响因子：
11.1
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA