Insulated transcriptional elements enable precise design of genetic circuits

Yeqing Zong, Haoqian M. Zhang, Cheng Lyu, Xiangyu Ji, Junran Hou, Xian Guo, Qi Ouyang () and Chunbo Lou ()
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Yeqing Zong: Chinese Academy of Sciences
Haoqian M. Zhang: Peking University
Cheng Lyu: Peking University
Xiangyu Ji: Chinese Academy of Sciences
Junran Hou: Chinese Academy of Sciences
Xian Guo: Chinese Academy of Sciences
Qi Ouyang: Peking University
Chunbo Lou: Chinese Academy of Sciences

Nature Communications, 2017, vol. 8, issue 1, 1-13

Abstract: Abstract 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 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.

Date: 2017
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DOI: 10.1038/s41467-017-00063-z

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