Rapid prototyping of biomolecular circuits through module characterization in cell-free expression systems

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Anton Frisk
Masters Thesis, Lund University

Over the past years, the field of synthetic biology has gained a significant array of tools and parts, making way for increasingly complex bio-molecular circuits to be constructed. The development of biocircuits can be facilitated by assembling parts in a less complex, cell-free, environment which contains only the machinery for gene transcription (TX) and translation (TL), which have been extracted from bacteria. In this project, a part library was collected and used to assemble DNA constructs for a newly designed biocircuit. An in vitro TX-TL extract was used to test the circuit modules using linear DNA, and in parallel with predictive modeling of the biomolecular reactions, the overall circuit design was evaluated. The results have given valuable insight into the performance of the circuit modules in a much shorter time than conventional in vivo cloning and testing would have achieved.