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3D structure of the long non-coding RNA Xist by complementary cryo-electron tomography and single particle cryo-electron microscopy

Objective

Recent studies have highlighted a pivotal role for long non-coding RNAs (lncRNAs) in normal physiology and disease. As a key example, Xist lncRNA is crucial during the early development of female mammals and is involved in many diseases, e.g. progeria and cancers. To date, cellular and developmental studies have advanced our knowledge of the function of Xist, but a detailed understanding of molecular mechanism is lacking.

Therefore, the overarching aim of my proposed research is to directly visualise Xist RNA/ ribonucleoprotein particles (RNPs) in cryo cells at sub-atomic resolution. To this end, I have formulated the following objectives: 1) preparing a cryo sample of mES cells expressing Xist RNA optimised for cryo electron tomography (ET); 2) visualising 3D-Xist RNPs in the mES cell; 3) generating homogenous pseudo-physiological Xist RNPs for single particle cryo-EM; 4) Solving the 3D sub-atomic resolution structure of Xist RNP; 5) Analysing a composite cryo-EM/-ET structure of the Xist RNPs in the mES cell.

To achieve this aim, I will employ an interdisciplinary approach including cutting-edge cryo-structural techniques and elegant mES cell biology. My supervisor Prof Brockdorff is a leading scientist in Xist lncRNA research and the host has a state-of-the-art cryo EM facility. The University of Oxford is a world-class research centre, providing excellent transferrable skills trainings and a fantastic infrastructure for career development.

The first high-resolution 3D structure of a lncRNA resulting from this innovative research will benefit not only the lncRNA research field, by providing novel molecular insight of the Xist lncRNA, but also the structural biology field, by expanding the 3D RNA structural potential. Dissemination and exploitation of this work, to be achieved by presenting in international conferences and publishing in high-impact open access journals, and communication, through public engagement, are an integral part of this proposed action.

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Funding Scheme

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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 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.

€ 183 454,80
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.

€ 183 454,80
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