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Development of chemical methods for DNA N6-methyladenine mapping

Objective

Herein, I propose to develop and adapt chemistry for the selective modification and tagging of N6-methyladenine (N6MeA) in the context of a DNA strand. Two promising chemical strategies will be applied, for which I have already established proofs-of-concept of efficiency and selectivity on DNA monomers (2'-deoxynucleosides and nucleotides).
Once such a specific chemical labelling protocol has been optimised, I will use it to map N6MeA in genomic DNA with two different approaches: 1) by chemical pulldown of N6-methylated DNA fragments, sequencing of the enriched fragments, and alignment to a reference genome to generate a low-resolution N6MeA map. 2) By analysing the influence of the introduced tags and modifications on the PCR outcome and take advantage of their stalling of polymerases.
The expected outcome is a first chemistry-assisted mapping of N6MeA. As for the chemical tagging of 5-formylcytosine (5fC) and 5-hydroxymethyluracil (5hmU), or oxidative and reductive bisulfite sequencing to sequence 5fC and 5-hydroxymethylcytosine (5hmC), all developed in the proposed host lab, it is expected to have a significant impact in the field. It will considerably facilitate the detection and mapping of genomic N6MeA in various species including mammals and humans, which is of tremendous importance to unravel the biological role of this DNA modification and identify novel biological as well as possible pathological pathways.

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MSCA-IF-EF-ST - Standard EF

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2017

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Coordinator

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 183 454,80
Address
TRINITY LANE THE OLD SCHOOLS
CB2 1TN CAMBRIDGE
United Kingdom

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Region
East of England East Anglia Cambridgeshire CC
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.

€ 183 454,80
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