Synthetic Biology – a heavy burden to bear?

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Rhys Algar, Guy-Bart Stan, Tom Ellis

Imperial College, United Kingdom

The expression of a synthetic gene circuit can have a detrimental effect on its chassis cell and these effects can feedback to the performance of the circuit. We investigate how a synthetic circuit uses cellular resources (eg. DNAP, RNAP, ribosomes, tRNA etc) to replicate and express. This is done by considering this shared ‘resource pool’ as an interface between the host cell and the synthetic circuit. We have developed a system for monitoring the availability of the resources used for protein expression within a cell (burden monitor). In order to test how different control points in heterologous gene expression affect the chassis cell, we have constructed a library of plasmid-based synthetic gene circuits with varying copy number, promoter strength, RBS strength and codon usage. By testing this library with our burden monitor we are able to uncover some unexpected behaviours and gain a greater understanding of how to optimise synthetic gene circuits to have minimal affect on their chassis cells.