合成生物学有可能彻底改变生物技术,公共卫生,和农业。最近的研究表明,植物作为合成生物学应用的底盘具有巨大的潜力。然而,仍然需要精确操纵植物生物生产代谢途径的工具。我们使用以配体特异性方式控制基因表达的细菌变构转录因子(aTFs),并测试了它们在植物中抑制半合成启动子的能力。我们还测试了它们对特定植物代谢物的抑制活性的调节,尤其是类苯丙烷相关分子.使用这些aTFs,我们还设计了能够计算布尔逻辑运算的合成遗传电路。三个aTF,Cour,FapR,还有TtgR,实现c.它们各自的靶启动子的95%抑制。对于TtgR,六倍去抑制可以通过诱导其配体积累来触发,显示其作为生物传感器的用途。此外,我们设计了使用AND的合成基因电路,NAND,imply,和NIMPLY布尔逻辑操作,并整合代谢物水平作为电路的输入。我们表明,生物传感器可以在植物中实施,以检测苯丙素类相关代谢物并激活遵循预定义逻辑的遗传电路,展示了它们作为控制植物代谢途径和促进天然产物生物生产的工具的潜力。
Synthetic biology has the potential to revolutionize biotechnology, public health, and agriculture. Recent studies have shown the enormous potential of plants as chassis for synthetic biology applications. However, tools to precisely manipulate metabolic pathways for bioproduction in plants are still needed. We used bacterial allosteric transcription factors (aTFs) that control gene expression in a ligand-specific manner and tested their ability to repress semi-synthetic promoters in plants. We also tested the modulation of their repression activity in response to specific plant metabolites, especially phenylpropanoid-related molecules. Using these aTFs, we also designed synthetic genetic circuits capable of computing Boolean logic operations. Three aTFs, CouR, FapR, and TtgR, achieved c. 95% repression of their respective target promoters. For TtgR, a sixfold de-repression could be triggered by inducing its ligand accumulation, showing its use as biosensor. Moreover, we designed synthetic genetic circuits that use AND, NAND, IMPLY, and NIMPLY Boolean logic operations and integrate metabolite levels as input to the circuit. We showed that biosensors can be implemented in plants to detect phenylpropanoid-related metabolites and activate a genetic circuit that follows a predefined logic, demonstrating their potential as tools for exerting control over plant metabolic pathways and facilitating the bioproduction of natural products.