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Content archived on 2024-06-18

Directed Evolution in vivo enabled through genetic circuits in a Synthetic Biology approach

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 (EuroSciVoc)

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Topic(s)

Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.

Call for proposal

Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.

FP7-PEOPLE-2013-IEF
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Funding Scheme

Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.

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

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Region
London Inner London — West Westminster
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

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