Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems.
Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems.
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DOI：
10.1038/s41467-020-20094-3
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发表时间：
2021-01-12
影响因子：
16.6
通讯作者：
Urtecho G
中科院分区：
文献类型：
--
作者：
Yu TC;Liu WL;Brinck MS;Davis JE;Shek J;Bower G;Einav T;Insigne KD;Phillips R;Kosuri S;Urtecho G
对生物系统进行工程设计的关键一步是精确调整遗传对环境刺激的反应的能力。在大肠杆菌可诱导启动子的情况下，我们对序列组成和基因表达之间的关系的不完全理解阻碍了我们预测地控制转录反应的能力。在这里，我们描述了8269个合理设计的、IPTG诱导的启动子的表达动态，这些启动子共同探索了RNA聚合酶和LacI抑制物结合位点强度的单独和组合效应。然后，我们将统计力学模型与测量的表达相匹配，该模型准确地模拟了基因的表达，并揭示了理论上最优的可诱导启动子的性质。此外，我们表征了三种不同的启动子结构，并表明启动子内结合位点的重新定位会影响启动子元件之间观察到的组合效应的类型。总之，这种方法使我们能够解构可诱导启动子元件之间的关系，并发现具有所需特征的工程可诱导启动子的实用见解。精确调整基因对环境刺激的反应是工程合成生物学系统的关键一步。在这里，作者描述了8269个IPTG诱导的启动子，以解构序列结构和基因表达之间的关系。
A crucial step towards engineering biological systems is the ability to precisely tune the genetic response to environmental stimuli. In the case of Escherichia coli inducible promoters, our incomplete understanding of the relationship between sequence composition and gene expression hinders our ability to predictably control transcriptional responses. Here, we profile the expression dynamics of 8269 rationally designed, IPTG-inducible promoters that collectively explore the individual and combinatorial effects of RNA polymerase and LacI repressor binding site strengths. We then fit a statistical mechanics model to measured expression that accurately models gene expression and reveals properties of theoretically optimal inducible promoters. Furthermore, we characterize three alternative promoter architectures and show that repositioning binding sites within promoters influences the types of combinatorial effects observed between promoter elements. In total, this approach enables us to deconstruct relationships between inducible promoter elements and discover practical insights for engineering inducible promoters with desirable characteristics. Precisely tuning the genetic response to environmental stimuli is a key step in engineering synthetic biology systems. Here, the authors profile 8269 IPTG-induced promoters to deconstruct the relationship between sequence architecture and gene expression.
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影响因子：
4.3
作者：
Brewster RC;Jones DL;Phillips R
通讯作者：
Phillips R
影响因子：
2
作者：
Boedicker, James Q.;Garcia, Hernan G.;Phillips, Rob
通讯作者：
Phillips, Rob
影响因子：
3.5
作者：
CHAN, B;BUSBY, S
通讯作者：
BUSBY, S
DOI：
10.1073/pnas.85.24.9683
发表时间：
1988-12-01
影响因子：
11.1
作者：
HABER, R;ADHYA, S
通讯作者：
ADHYA, S
影响因子：
3.2
作者：
Djordjevic, Marko
通讯作者：
Djordjevic, Marko