Cell-Free Gene Expression (CFE) Systems

Cell-free gene expression (CFE) systems provide a bottom-up approach for synthetic biology engineering. As an introduction, CFE systems require a cell extract, an RNA/DNA template and an metabolite solution, to make mRNA and proteins within seconds to minutes timescales.

CFE systems are actually quite complicated systems. While they are devoid of a cell membrane, cell wall and genomic DNA, the cell-free extracts contain approximately 400-1000 distinct proteins (estimated from high-resolution mass spectrometry), a multitude of RNA and metabolites species, and a myriad of underlying protein-metabolite and protein-protein interactions that are crucial to the biological activity of CFE systems. All of this still requires further characterisation through omics technologies. We are generally interested in being able to quantify and standardise these processes in line with the underlying principles of synthetic biology, as well harnessing the distinct properties each extracts provides. For example, while Escherichia coli is the dominant CFE system, we are also interested in exploring the potential of developing non-model CFE systems for different applications. We have developed a range of CFE systems, with the expertise, protocols, extracts and plasmid expression systems for the following bacteria.

• Escherichia coli (BL21, Rosetta, MG1655)

• Bacillus megaterium - Protein production and antimicrobial testing applications

• Streptomyces spp., including Streptomyces venezuelae - Natural product biosynthesis applications

• Klebsiella pneumoniae - A model CFE system for studying antimicrobials and resistance (in review)

• New systems (Gram-negative, Gram-positive) in development

We collaborate with a range of academic groups and industry both from within the UK and internationally.

• For expressions of interest, feel free to email Dr Simon Moore at simon.moore[at]qmul.ac.uk