Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Nat Commun. 2020 Nov 10;11(1):5690. doi: 10.1038/s41467-020-19126-9.
Synthetic biology has the potential to bring forth advanced genetic devices for applications in healthcare and biotechnology. However, accurately predicting the behavior of engineered genetic devices remains difficult due to lack of modularity, wherein a device's output does not depend only on its intended inputs but also on its context. One contributor to lack of modularity is loading of transcriptional and translational resources, which can induce coupling among otherwise independently-regulated genes. Here, we quantify the effects of resource loading in engineered mammalian genetic systems and develop an endoribonuclease-based feedforward controller that can adapt the expression level of a gene of interest to significant resource loading in mammalian cells. Near-perfect adaptation to resource loads is facilitated by high production and catalytic rates of the endoribonuclease. Our design is portable across cell lines and enables predictable tuning of controller function. Ultimately, our controller is a general-purpose device for predictable, robust, and context-independent control of gene expression.
合成生物学有可能带来先进的遗传设备,应用于医疗保健和生物技术领域。然而,由于缺乏模块化,设计的遗传设备的行为难以准确预测,其中设备的输出不仅取决于预期的输入,还取决于其上下文。缺乏模块化的一个原因是转录和翻译资源的加载,这会导致原本独立调控的基因之间发生耦合。在这里,我们量化了工程化哺乳动物遗传系统中资源加载的影响,并开发了一种基于内切核糖核酸酶的前馈控制器,可以将感兴趣的基因的表达水平自适应地调整到哺乳动物细胞中的显著资源加载。内切核糖核酸酶的高生产和催化速率促进了对资源负载的近乎完美的适应。我们的设计可在细胞系之间移植,并能够对控制器功能进行可预测的调整。最终,我们的控制器是一种通用设备,用于可预测、稳健和独立于上下文的基因表达控制。