Lane Center for Computational Biology, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.
Nat Nanotechnol. 2013 Aug;8(8):602-8. doi: 10.1038/nnano.2013.132. Epub 2013 Jul 14.
The integration of synthetic and cell-free biology has made tremendous strides towards creating artificial cellular nanosystems using concepts from solution-based chemistry, where only the concentrations of reacting species modulate gene expression rates. However, it is known that macromolecular crowding, a key feature in natural cells, can dramatically influence biochemical kinetics via volume exclusion effects, which reduce diffusion rates and enhance binding rates of macromolecules. Here, we demonstrate that macromolecular crowding can increase the robustness of gene expression by integrating synthetic cellular components of biological circuits and artificial cellular nanosystems. Furthermore, we reveal how ubiquitous cellular modules, including genetic components, a negative feedback loop and the size of the crowding molecules can fine-tune gene circuit response to molecular crowding. By bridging a key gap between artificial and living cells, our work has implications for efficient and robust control of both synthetic and natural cellular circuits.
合成生物学和无细胞生物学的融合在利用基于溶液化学的概念来创建人工细胞纳米系统方面取得了巨大进展，在基于溶液化学的概念中，只有反应物种的浓度调节基因表达率。然而，众所周知，生物细胞的一个关键特征——大分子拥挤，通过体积排除效应可以显著影响生化动力学，从而降低扩散率并增强大分子的结合率。在这里，我们通过整合生物电路和人工细胞纳米系统的合成细胞成分来证明，大分子拥挤可以提高基因表达的稳健性。此外，我们揭示了包括遗传成分、负反馈回路和拥挤分子的大小在内的普遍存在的细胞模块如何微调基因电路对分子拥挤的反应。通过弥合人工细胞和活细胞之间的关键差距，我们的工作对于有效和稳健地控制合成和自然细胞电路都具有重要意义。
