Objective Given our limited ability to rationally design proteins to our needs, their supply for biotechnological solutions leading to a sustainable Bioeconomy is relying on proteins found in nature and methods for deriving improved versions. Proteins evolved their structure to fulfil certain functions in very specific niches, but these do not resemble optimal properties in the new context of biotechnological applications. Hence, improving the activity, affinity and stability of a protein is key, but technologically challenging. The current methods for improving proteins through directed evolution require intense human intervention and are limited in their ability to sample sequence space beyond single and double mutations. This project will establish a novel system, in which the whole process of mutation and selection takes place in vivo inside an E. coli host, and can be controlled through a genetic network designed using a Synthetic Biology approach. It will allow for the exploration of a larger sequence space during the evolutionary process, while controlling mutagenesis rate and selection stringency. Our preliminary data demonstrates a groundbreaking novel method to target mutagenesis in vivo. I will extend its mutation diversity by recruiting engineered error prone polymerases to the site of the targeted DNA damage. Using a SynBio approach, I will develop a control circuitry, which will stop the targeted mutagenesis activity, once a variant fulfils the selection criteria. While protecting the identity of the selected sequence, it will mark the successful cell by expression of a fluorescent reporter for selection. In a prototype application I will generate variants of the Quorum Sensing transcription factor, LuxR, with high affinities to different signalling molecules – a potential toolset for orthogonal cellular signalling applications in industrially relevant co-cultures. The resulting directed evolution system will be readily adaptable to different proteins. Fields of science natural sciencesbiological sciencessynthetic biologynatural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesevolutionary biologynatural sciencesbiological sciencesgeneticsmutation Programme(s) FP7-PEOPLE - Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Topic(s) FP7-PEOPLE-2013-IEF - Marie-Curie Action: "Intra-European fellowships for career development" Call for proposal FP7-PEOPLE-2013-IEF See other projects for this call Funding Scheme MC-IEF - Intra-European Fellowships (IEF) Coordinator IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE EU contribution € 221 606,40 Address SOUTH KENSINGTON CAMPUS EXHIBITION ROAD SW7 2AZ LONDON United Kingdom See on map Region London Inner London — West Westminster Activity type Higher or Secondary Education Establishments Administrative Contact Brooke Alasya (Ms.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data
