Construction of integrated gene logic-chip
Construction of integrated gene logic-chip
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DOI：
10.1038/s41565-018-0202-3
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发表时间：
2018-10-01
影响因子：
38.3
通讯作者：
Tadakuma, Hisashi
中科院分区：
文献类型：
--
作者：
Masubuchi, Takeya;Endo, Masayuki;Tadakuma, Hisashi
在合成生物学中，基因表达的控制需要对生物信号进行多步骤处理。关键步骤是感知环境、计算信息和输出产品(1)。为了实现这些功能，需要酶和底物基因的费力的组合网络，并且为了解决问题，已经引入了复杂的设计自动化工具(2)。然而，遗传电路的复杂性仍然很低，因为很难完全避免电路之间的串扰。在这里，我们通过将执行器和传感器集成到基于 DNA 折纸的纳米芯片上，制作了一个正交独立设备，该纳米芯片包含酶、T7 RNA 聚合酶 (RNAP) 和多个靶基因底物。这种基因纳米芯片正交转录自己的基因，DNA折纸的纳米布局能力使我们能够通过控制酶与目标基因之间的分子间距离来合理设计基因表达水平。我们进一步集成了可重编程逻辑门，使纳米芯片能够响应油包水液滴并计算它们的小RNA (miRNA)图谱，这表明纳米芯片可以充当基因逻辑芯片。我们在纳米芯片上集成组件的方法可能为大规模集成遗传电路提供基础。
In synthetic biology, the control of gene expression requires a multistep processing of biological signals. The key steps are sensing the environment, computing information and outputting products(1). To achieve such functions, the laborious, combinational networking of enzymes and substrate-genes is required, and to resolve problems, sophisticated design automation tools have been introduced(2). However, the complexity of genetic circuits remains low because it is difficult to completely avoid crosstalk between the circuits. Here, we have made an orthogonal self-contained device by integrating an actuator and sensors onto a DNA origami-based nanochip that contains an enzyme, T7 RNA polymerase (RNAP) and multiple target-gene substrates. This gene nanochip orthogonally transcribes its own genes, and the nano-layout ability of DNA origami allows us to rationally design gene expression levels by controlling the intermolecular distances between the enzyme and the target genes. We further integrated reprogrammable logic gates so that the nanochip responds to water-in-oil droplets and computes their small RNA (miRNA) profiles, which demonstrates that the nanochip can function as a gene logic-chip. Our approach to component integration on a nanochip may provide a basis for large-scale, integrated genetic circuits.