Lawrence Berkeley National Laboratory
Programming mRNA decay to modulate synthetic circuit resource allocation.
- Author(s): Venturelli, Ophelia S
- Tei, Mika
- Bauer, Stefan
- Chan, Leanne Jade G
- Petzold, Christopher J
- Arkin, Adam P
- et al.
Published Web Locationhttps://doi.org/10.1038/ncomms15128
Synthetic circuits embedded in host cells compete with cellular processes for limited intracellular resources. Here we show how funnelling of cellular resources, after global transcriptome degradation by the sequence-dependent endoribonuclease MazF, to a synthetic circuit can increase production. Target genes are protected from MazF activity by recoding the gene sequence to eliminate recognition sites, while preserving the amino acid sequence. The expression of a protected fluorescent reporter and flux of a high-value metabolite are significantly enhanced using this genome-scale control strategy. Proteomics measurements discover a host factor in need of protection to improve resource redistribution activity. A computational model demonstrates that the MazF mRNA-decay feedback loop enables proportional control of MazF in an optimal operating regime. Transcriptional profiling of MazF-induced cells elucidates the dynamic shifts in transcript abundance and discovers regulatory design elements. Altogether, our results suggest that manipulation of cellular resource allocation is a key control parameter for synthetic circuit design.