Engineering cell-free systems for synthetic biologists

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Sheahan, Taylor Anne
University of Lethbridge. Faculty of Arts and Science
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Lethbridge, Alta. : University of Lethbridge, Dept. of Chemistry and Biochemistry
Synthetic biology (synbio) has emerged as a transformative scientific field with immense potential to address a wide-range of global problems. A specific sub-field of synbio utilizes cellular biomolecular machinery outside of a living cell. In theory, these “cell-free” systems offer a simpler approach and unique features compared to cell-based systems for biotechnology development. However, in practice limited accessibility and poor protein synthesis capacity hinder the overall scope and application of cell-free synbio. To address these challenges, it was our goal to create new engineering tools that will help expand the overall utility of cell-free expression systems. Data presented here provides: 1) detailed methods for the in-house preparation of a cost-effective in vitro reconstituted cell-free system, 2) an in-depth proteomic analysis of the system building blocks as a tool to characterize the composition and inform optimization, and 3) an improvement to protein synthesis capacity by modifying the ribosome composition. Furthermore, a critical assessment of the regulatory landscape is provided, promoting the safe and responsible use of cell-free synbio.
synthetic biology , cell-free systems , Synthetic biology , Proteins--Synthesis , Proteomics , Biotechnology , Bioengineering , Dissertations, Academic