Biological 2-input decoder circuit in human cells.

Biological 2-input decoder circuit in human cells.
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DOI:
10.1021/sb4001596
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发表时间:
2014-08-15
影响因子:
4.7
通讯作者:
Bleris, Leonidas
Bleris, Leonidas
中科院分区:
生物学2区
文献类型:
--
作者:
Guinn, Michael;Bleris, Leonidas

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解码器是将信息从n个输入转换为最多2n个输出的组合电路。这一操作在计算系统中非常重要,但在合成生物学中尚未得到充分的探索。在这里,我们提出了一个合成基因网络架构,作为人类细胞中的生物解码器,将2个输入转换为4个输出。作为原理证明,我们使用小分子模拟两个输入和荧光报告作为相应的四个输出。实验采用瞬时转染人肾胚胎细胞,并通过荧光显微镜和流式细胞术进行表征。我们显示了一个明确的分离之间的开和关平均荧光强度状态。此外,我们采用积分平均荧光强度来表征电路,并表明与平均荧光强度相比,该指标对转染条件更具鲁棒性。总之,我们提出了遗传解码器的第一个实现。这种组合系统对于工程高阶电路以及容纳具有内源性细胞功能的多路接口具有价值。
Decoders are combinational circuits that convert information from n inputs to a maximum of 2n outputs. This operation is of major importance in computing systems yet it is vastly underexplored in synthetic biology. Here, we present a synthetic gene network architecture that operates as a biological decoder in human cells, converting 2 inputs to 4 outputs. As a proof-of-principle, we use small molecules to emulate the two inputs and fluorescent reporters as the corresponding four outputs. The experiments are performed using transient transfections in human kidney embryonic cells and the characterization by fluorescence microscopy and flow cytometry. We show a clear separation between the ON and OFF mean fluorescent intensity states. Additionally, we adopt the integrated mean fluorescence intensity for the characterization of the circuit and show that this metric is more robust to transfection conditions when compared to the mean fluorescent intensity. To conclude, we present the first implementation of a genetic decoder. This combinational system can be valuable toward engineering higher-order circuits as well as accommodate a multiplexed interface with endogenous cellular functions.
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