Characterisation of genetic control elements for synthetic biology using the Spinach RNA aptamer

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Georgios Pothoulakis, Tom Ellis

Imperial College London, United Kingdom

Accurate characterisation of genetic control elements is essential for the predictable engineering of synthetic biology systems. Real-time evaluation of biological parts like promoters, riboswitches and ribosome binding sites enables design of customised gene expression and the balancing metabolic pathways. The current standard for in vivo characterisation of control elements is through the use of fluorescent reporter proteins such as GFP. However, gene expression is a multi-step process, specifically with RNA production as a key intermediate. The ability of the Spinach RNA aptamer to mimic the characteristics of the GFP protein; bypasses several disadvantages and limitations of older RNA measurement methods and provides information about the translational efficiency and transcriptional strength of several control elements when the aptamer is used in the context of an ORF.