Amplification of small molecule-inducible gene expression via tuning of intracellular receptor densities.
Amplification of small molecule-inducible gene expression via tuning of intracellular receptor densities.
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DOI：
10.1093/nar/gku1388
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发表时间：
2015-02-18
影响因子：
14.9
通讯作者：
Buck M
中科院分区：
文献类型：
--
作者：
Wang B;Barahona M;Buck M
原核生物中的配体应答转录因子发现了简单的小分子诱导基因表达系统。这些已被广泛用于调节蛋白质生产和精细化学品的相关生物合成。然而，传统上使用的启动子和蛋白质工程方法通常对可预测地和快速地调节诱导型表达系统的表达谱造成显著限制。在这里，我们提出了一个新的统一和合理的调整方法，以扩大灵敏度和多功能小分子诱导表达系统的动态范围。我们采用细胞内受体浓度的系统变化进行转录控制。我们表明，低密度的阻遏物受体（例如TetR和ArsR）在细胞中可以显着增加的灵敏度和动态范围，而高激活受体（例如LuxR）密度达到相同的结果。受体的细胞内浓度可以通过调节其同源驱动启动子的强度以离散和连续模式调节。我们在几个合成受体介导的传感电路中举例说明了这种方法，包括可调的基于细胞的砷传感器。该方法提供了一种新的范式，可预测地调整和扩增配体响应基因表达，在合成生物学和工业生物技术中具有潜在的应用。
Ligand-responsive transcription factors in prokaryotes found simple small molecule-inducible gene expression systems. These have been extensively used for regulated protein production and associated biosynthesis of fine chemicals. However, the promoter and protein engineering approaches traditionally used often pose significant restrictions to predictably and rapidly tune the expression profiles of inducible expression systems. Here, we present a new unified and rational tuning method to amplify the sensitivity and dynamic ranges of versatile small molecule-inducible expression systems. We employ a systematic variation of the concentration of intracellular receptors for transcriptional control. We show that a low density of the repressor receptor (e.g. TetR and ArsR) in the cell can significantly increase the sensitivity and dynamic range, whereas a high activator receptor (e.g. LuxR) density achieves the same outcome. The intracellular concentration of receptors can be tuned in both discrete and continuous modes by adjusting the strength of their cognate driving promoters. We exemplified this approach in several synthetic receptor-mediated sensing circuits, including a tunable cell-based arsenic sensor. The approach offers a new paradigm to predictably tune and amplify ligand-responsive gene expression with potential applications in synthetic biology and industrial biotechnology.
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影响因子：
9.9
作者：
Cox, Robert Sidney 3rd;Surette, Michael G;Elowitz, Michael B
通讯作者：
Elowitz, Michael B
影响因子：
14.9
作者：
Mazumder M;McMillen DR
通讯作者：
McMillen DR
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.2
作者：
DIORIO, C;CAI, J;DUBOW, MS
通讯作者：
DUBOW, MS
DOI：
10.1099/mic.0.067975-0
发表时间：
2013-07
期刊：
Microbiology (Reading, England)
影响因子：
--
作者：
Arpino JAJ;Hancock EJ;Anderson J;Barahona M;Stan GV;Papachristodoulou A;Polizzi K
通讯作者：
Polizzi K