Insulated transcriptional elements enable precise design of genetic circuits.
Insulated transcriptional elements enable precise design of genetic circuits.
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绝缘转录元件可实现遗传电路的精确设计
DOI：
10.1038/s41467-017-00063-z
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发表时间：
2017-07-03
影响因子：
16.6
通讯作者：
Lou C
中科院分区：
文献类型：
--
作者：
Zong Y;Zhang HM;Lyu C;Ji X;Hou J;Guo X;Ouyang Q;Lou C
生物系统的合理工程通常因细胞成分之间在多个层面上复杂但不必要的相互作用而变得复杂。在这里，我们在原核转录水平上解决这个问题，通过隔离最小的启动子和操纵子以防止它们的相互作用，并在没有自由参数的情况下实现合成转录的生物物理建模。这种方法允许遗传电路设计具有非凡的精度和多样性，从而将电路工程的设计-构建-测试-学习周期简化为混合匹配工作流程。作为演示，使用我们的绝缘转录元件从头开始设计编码非门功能的组合启动子，平均误差<1.5倍，成功率>96%。此外，无需任何试错工作即可获得具有执行条带形成功能的不相干前馈循环的四节点转录网络。这种基于绝缘的工程策略提高了遗传电路技术的分辨率，并为系统和合成生物学设计遗传电路提供了一种简单的方法。
Rational engineering of biological systems is often complicated by the complex but unwanted interactions between cellular components at multiple levels. Here we address this issue at the level of prokaryotic transcription by insulating minimal promoters and operators to prevent their interaction and enable the biophysical modeling of synthetic transcription without free parameters. This approach allows genetic circuit design with extraordinary precision and diversity, and consequently simplifies the design-build-test-learn cycle of circuit engineering to a mix-and-match workflow. As a demonstration, combinatorial promoters encoding NOT-gate functions were designed from scratch with mean errors of <1.5-fold and a success rate of >96% using our insulated transcription elements. Furthermore, four-node transcriptional networks with incoherent feed-forward loops that execute stripe-forming functions were obtained without any trial-and-error work. This insulation-based engineering strategy improves the resolution of genetic circuit technology and provides a simple approach for designing genetic circuits for systems and synthetic biology.