Zymogens and Transmembrane Activation of Transcription in Synthetic Cells
Andersen, D. G., Pedersen, A. B., Jørgensen, M. H., Montasell, M. C., Søgaard, A. B., Chen, G., Schroeder, A., Andersen, G. R., Zelikin, A. N., Zymogens and Transmembrane Activation of Transcription in Synthetic Cells. Adv. Mater. 2023, 2309385.
In this work, we engineer synthetic cells equipped with an artificial signaling pathway that connects an extracellular trigger event to the activation of intracellular transcription. Learning from nature, we do so via an engineering of responsive enzymes, such that activation of enzymatic activity can be triggered by an external biochemical stimulus. We engineer reversibly deactivated creatine kinase to achieve triggered production of ATP, and a reversibly deactivated nucleic acid polymerase for on-demand synthesis of RNA. We also design an extracellular, enzyme-activated production of a diffusible zymogen activator. The key achievement of our work is that we illustrate the importance of cellularity whereby the separation of biochemical partners is essential to resolve their incompatibility, to enable transcription within the confines of a synthetic cell. The herein designed biochemical pathway and the engineered synthetic cells are arguably primitive compared to their natural counterpart. Nevertheless, our results present a significant step towards the design of synthetic cells with responsive behaviour, en route from abiotic to life-like cell mimics.