A portable expression resource for engineering cross-species genetic circuits and pathways.
A portable expression resource for engineering cross-species genetic circuits and pathways.
复制标题
DOI：
10.1038/ncomms8832
复制
发表时间：
2015-07-17
影响因子：
16.6
通讯作者：
Salis HM
中科院分区：
文献类型：
--
作者：
Kushwaha M;Salis HM
可以重新设计遗传回路和代谢途径，使生物体能够处理信号并制造有用的化学物质。然而，它们的功能目前依赖于生物体特异性的调节部分，将合成生物学和代谢工程分割成宿主特异性的领域。为了统一努力，我们在这里设计了一种跨物种表达资源，使电路和通路能够重复使用相同的遗传部分，同时在不同的生物体中发挥相似的作用。我们的工程系统结合了混合反馈控制环和跨物种翻译信号，可以在没有宿主特异性启动子的情况下自主调节正交聚合酶的表达，从而在多种革兰氏阳性和革兰氏阴性细菌中实现无毒且可调节的基因表达。将 50 个特征系统变体与机械建模相结合，我们展示了跨物种表达资源的动态、容量和毒性如何通过控制环路的架构和反馈强度进行控制。我们还通过重复使用相同的遗传部分在模型和非模型宿主中表达生物合成途径来展示该资源的一种应用。 生物体特异性的遗传部分通常用于表达电路和通路，限制了它们的可移植性。在这里，作者设计了一种跨物种表达资源，无需使用宿主特异性部分，即可控制非模型细菌中的蛋白质和途径表达。
Genetic circuits and metabolic pathways can be reengineered to allow organisms to process signals and manufacture useful chemicals. However, their functions currently rely on organism-specific regulatory parts, fragmenting synthetic biology and metabolic engineering into host-specific domains. To unify efforts, here we have engineered a cross-species expression resource that enables circuits and pathways to reuse the same genetic parts, while functioning similarly across diverse organisms. Our engineered system combines mixed feedback control loops and cross-species translation signals to autonomously self-regulate expression of an orthogonal polymerase without host-specific promoters, achieving nontoxic and tuneable gene expression in diverse Gram-positive and Gram-negative bacteria. Combining 50 characterized system variants with mechanistic modelling, we show how the cross-species expression resource's dynamics, capacity and toxicity are controlled by the control loops' architecture and feedback strengths. We also demonstrate one application of the resource by reusing the same genetic parts to express a biosynthesis pathway in both model and non-model hosts. Organism-specific genetic parts are often used to express circuits and pathways, limiting their portability. Here the authors engineer a cross-species expression resource, without using host-specific parts, to control protein and pathway expression in non-model bacteria.