Objective
We propose to investigate the enzymes responsible for DNA replication and repair in micromanipulation experiments with a resolution of a single base. The detailed mechanism by which DNA is synthesized base after base and the coordination of the enzymes involved in this process are not fully understood. We shall develop new magnetic tweezers using lithographic techniques associated with evanescent field detection to address these issues. We shall build arrays of these devices working in parallel, each one on a single DNA molecule and where the measurement of its extension reveals enzymatic activity. The DNA molecule in these devices will form a hairpin the opening of which can be detected with a single base resolution.
We will study the different enzymes involved in DNA replication. Firstly, we wish to follow in real time the incorporation of bases one by one by a DNA-polymerase and investigate the proof-reading mechanism of this enzyme. We shall also investigate the translocation mechanisms of different helicases involved in DNA replication and repair. Finally, we plan to study the cooperative action between different enzymes involved in the replication machinery with the help of parallelized micro-tweezers: the coupling between helicase and primase in the lagging strand synthesis, the coupling between the helicase and polymerase during leading strand synthesis and the coordination between leading and lagging strand synthesis.
Moreover observing a DNA-polymerase at the single base level is the first step of a DNA sequencing method. Preliminary experiments demonstrate that the unzipping assay is a new way to determine the position of a small DNA sequence with single base resolution. We shall investigate different experimental schemes to achieve this goal.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: https://op.europa.eu/en/web/eu-vocabularies/euroscivoc.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: https://op.europa.eu/en/web/eu-vocabularies/euroscivoc.
- natural sciences biological sciences genetics DNA
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
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.
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
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Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
ERC-2010-AdG_20100224
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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.
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.
Host institution
75794 PARIS
France
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.