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Imaging nucleic acid metabolism in cells

Objective

In this project I, Pawan Kumar, a Nucleic Acid Chemist seek to team up with a pioneer in Nucleic Acid Chemistry and Biology (Professor Tom Brown) to carry out a highly interdisciplinary study at the University of Oxford to obtain deeper insights into DNA and RNA synthesis and metabolism in living cells and its application to cancer research. I propose to detect and quantify nucleic acid synthesis in proliferating cells by fluorescence without the requirement for toxic metal ions or antibodies. Earlier known methods such as [3H]thymidine and 5-bromo-2'-deoxyuridine labelling are either slow and labour intensive or require the use of harsh conditions. Incorporation of 5-ethynyl-2'-deoxyuridine into newly synthesized DNA and its subsequent detection with an azide derivative of a fluorescent dye under copper catalyzed alkyne azide cycloaddition (CuAAC) reaction conditions presents a better alternative. However, cytotoxicity of copper salts restricts its use for living cells. I will use the strain promoted alkyne azide cycloaddition reaction, Diels-Alder reaction, and inverse electron demand Diels-Alder reaction to study cellular DNA and RNA. None of these reactions require the use of toxic metal salts. I will develop the conditions under which both DNA and RNA will be stable, so that it will be possible to isolate intact fluorescent nucleic acids from cells for detailed analysis. I will prepare the modified nucleosides and use them to label newly synthesized DNA and RNA in cells enabling their detection by reaction with fluorophores by using metal free click ligation reactions. The study will provide a better understanding of the mechanisms regulating DNA replication and the interplay between transcription and DNA replication. In the project I will develop techniques to provide information on the toxic effect of antimetabolites used commonly in anti-cancer therapies, and for identifying the mechanisms of viral replication and understanding the viral life cycle.

Fields of science (EuroSciVoc)

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Programme(s)

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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.

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.

MSCA-IF-EF-ST - Standard EF

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Call for proposal

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

(opens in new window) H2020-MSCA-IF-2014

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Coordinator

THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
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.

€ 195 454,80
Address
WELLINGTON SQUARE UNIVERSITY OFFICES
OX1 2JD Oxford
United Kingdom

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Region
South East (England) Berkshire, Buckinghamshire and Oxfordshire Oxfordshire
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.

€ 195 454,80
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