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The behaviour of engineered biological systems can be subject to deviation from design due to unpredictable changes in growth conditions, host response and metabolic load. Control engineering approaches are key to automatically maintain robust performance of designed systems despite such variations. RNA-controlled gene expression creates new possibilities for systematic, fast-responding and resource-efficient implementations of synthetic biology systems in living cells. Here, we demonstrate that a rational, modular design framework can provide an extensible platform for m odular A rtificial R NA i nterference (mARi) for robust, portable and multiplexed post-transcriptional regulation in Escherichia coli. We used mARi to construct a host-aware negative feedback control system that regulated protein production in response to cellular stress. We demonstrated that this mARi-based feedback control system resulted in an improvement in protein production up to 5-fold and was robust to perturbations in temperature, medium, scale, cell strain and gene product without further tuning. We anticipate that the use of robust RNA-based genetic control within an extensible DNA assembly framework, such as mARi, will have widespread applications in the realisation of sophisticated engineered biological systems, where dynamic regulation can provide improvements in gene circuit stability and microbial production. |
