Mahajan Tarun, Rai Kshitij
Department of Electrical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi, India.
PLoS One. 2018 Feb 1;13(2):e0183242. doi: 10.1371/journal.pone.0183242. eCollection 2018.
Synthetic biology has enabled the creation of biological reconfigurable circuits, which perform multiple functions monopolizing a single biological machine; Such a system can switch between different behaviours in response to environmental cues. Previous work has demonstrated switchable dynamical behaviour employing reconfigurable logic gate genetic networks. Here we describe a computational framework for reconfigurable circuits in E.coli using combinations of logic gates, and also propose the biological implementation. The proposed system is an oscillator that can exhibit tunability of frequency and amplitude of oscillations. Further, the frequency of operation can be changed optogenetically. Insilico analysis revealed that two-component light systems, in response to light within a frequency range, can be used for modulating the frequency of the oscillator or stopping the oscillations altogether. Computational modelling reveals that mixing two colonies of E.coli oscillating at different frequencies generates spatial beat patterns. Further, we show that these oscillations more robustly respond to input perturbations compared to the base oscillator, to which the proposed oscillator is a modification. Compared to the base oscillator, the proposed system shows faster synchronization in a colony of cells for a larger region of the parameter space. Additionally, the proposed oscillator also exhibits lesser synchronization error in the transient period after input perturbations. This provides a strong basis for the construction of synthetic reconfigurable circuits in bacteria and other organisms, which can be scaled up to perform functions in the field of time dependent drug delivery with tunable dosages, and sets the stage for further development of circuits with synchronized population level behaviour.
合成生物学已能够创建生物可重构电路,这种电路执行多种功能并独占单个生物机器;这样的系统可以根据环境线索在不同行为之间切换。先前的工作已经证明了利用可重构逻辑门遗传网络实现的可切换动态行为。在此,我们描述了一种使用逻辑门组合构建大肠杆菌中可重构电路的计算框架,并提出了其生物学实现方式。所提出的系统是一个振荡器,它能够展现出振荡频率和幅度的可调性。此外,其工作频率可以通过光遗传学方法改变。计算机模拟分析表明,双组分光系统在一定频率范围内对光作出响应,可用于调节振荡器的频率或完全停止振荡。计算建模显示,将两个以不同频率振荡的大肠杆菌菌落混合会产生空间拍频模式。此外,我们表明,与基础振荡器相比,所提出的振荡器对输入扰动的响应更强健,基础振荡器是所提出振荡器的改进对象。与基础振荡器相比,所提出系统在更大参数空间区域内的细胞群体中表现出更快的同步性。此外,所提出的振荡器在输入扰动后的瞬态期也表现出较小的同步误差。这为在细菌和其他生物体中构建合成可重构电路提供了有力基础,这些电路可扩大规模以在时间依赖性药物递送领域中实现可调剂量功能,并为具有同步群体水平行为的电路进一步发展奠定了基础。