14TSB_SynBio A High Throughput Miniaturised Mass Spectrometry Tool for Profiling Synthetic Design Libraries

Over the last 10 years, the new field of synthetic biology has advanced the existing science of genetic modification by applying principles of engineering; design, simulation and testing. This approach has allowed us to predictably create exciting new technologies by modifying safe microbes with customised DNA to perform new tasks including computation, multi-input environmental sensing and efficient production of medicinal drugs and bioenergy, primarily through metabolic pathway engineering. As our ability to write DNA and engineer microbes accelerates, we are becoming limited by the lack of high-data, high-throughput cell measurement methods available with which to assess the performance of customised strains. The project links foundational synthetic biology and DNA assembly expertise at Imperial College London with microfluidics and mass spectrometry expertise at the industrial partner, Sphere Fluidics Limited, in order to build, test and demonstrate a novel microfluidics-mass spectrometry tool that should allow in-depth measurement of the performance of thousands of engineered microbes per run. The intention of the project is to develop an equipment set-up (interfaced microfluidics and mass spectrometry) and a methodology that together is a platform tool with which to characterise collections of thousands of engineered microbe strains. This tool can then be sold downstream as a service (or as an equipment set-up) for companies and institutions involved in synthetic biology.

Over the last 10 years, the new field of synthetic biology has advanced the existing science of genetic modification by applying principles of engineering; design, simulation and testing. This approach has allowed us to predictably create exciting new technologies by modifying safe microbes with customised DNA to perform new tasks including computation, multi-input environmental sensing and efficient production of medicinal drugs and bioenergy, primarily through metabolic pathway engineering. As our ability to write DNA and engineer microbes accelerates, we are becoming limited by the lack of high-data, high-throughput cell measurement methods available with which to assess the performance of customised strains. The project links foundational synthetic biology and DNA assembly expertise at Imperial College London with microfluidics and mass spectrometry expertise at the industrial partner, Sphere Fluidics Limited, in order to build, test and demonstrate a novel microfluidics-mass spectrometry tool that should allow in-depth measurement of the performance of thousands of engineered microbes per run. The intention of the project is to develop an equipment set-up (interfaced microfluidics and mass spectrometry) and a methodology that together is a platform tool with which to characterise collections of thousands of engineered microbe strains. This tool can then be sold downstream as a service (or as an equipment set-up) for companies and institutions involved in synthetic biology.

This decade, synthetic biology is expected to move from the demonstration of new technologies to being a major part of applied bioscience research and commercial biotechnology. Already, the synthetic biology market was worth £1.9 billion in 2013 and is expected to grow to around £10.6 billion by 2018 (Transparency Market Research (2012): Synthetic Biology Market, Global Industry Analysis, Size, Growth, Share and Forecast, 2012-2018). Within this market, there is an increasing need for new enabling technologies to help drive the research and development of future products.
The project proposed here intends to have a major impact on the global synthetic biology sector, in both the commercial market and in the rapidly-expanding academic research field, by providing a new enabling technology as a tool/service for screening and characterising engineered strains. This short project brings commercial microfluidics and mass spectrometry expertise together with academic excellence in synthetic biology to transform the way engineered microbial strains are screened and characterised when constructed using combinatorial design methods that yield construct libraries. Traditionally, screening of combinatorial libraries has been limited to engineered strains that incorporate fluorescence output into their design, radically limiting what products can be quickly screened-for. By bringing together microfluidics, mass spectrometry and synthetic biology, this project will develop a new enabling technology that does not require fluorescence to assess and sort thousands of engineered strains. This will impact synthetic biology and especially metabolic engineering projects, by opening-up new avenues where combinatorial designs can be tested without use of fluorescent proteins. Potentially, this could catalyse a step-change in the productivity of synthetic biology research and development, as design limitations will be significantly reduced. Further impact will be afforded by the greater amount of data that mass spectrometry produces per characterisation. Measurement by mass spectrometry reveals not only the quantity and quality of the product of an engineered strain but also provides a wealth of information on the health of the engineered strain, such as the energy availability (e.g. ATP/ADP ratios). These secondary data will be invaluable in selecting the most efficient engineered strains and will greatly inform future designs. The impact of switching from fluorescence-based characterisation to mass spectrometry-based characterisation has the potential to turn synthetic biology into a 'big data' science, where computation and prediction are enabled by rich datasets. Doing this using microfluidics also guarantees low-cost and affordability, making the technology developed in this project more likely to impact on a wider audience, both in academia and in industry.
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