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A DUAL gRNA system for functional assessment of ENhancers in Pluripotency

Objective

Enhancers are cis-regulatory genomic regions that can modulate gene expression in a cell type-specific and time-controlled manner to regulate cellular behaviour. ChIP-seq and RNA-seq technologies have allowed genome-wide identification of potential enhancers based on the correlation among specific chromatin marks, transcription factor biding sites and gene expression. However, strategies to validate their function are still limited. Recently, it has been demonstrated that the CRISPR/Cas9 genome editing technology can be used to delete large genomic regions at high frequency by co-transfection of two guide RNAs (gRNAs) targeting collinear genomic sites and subsequent NHEJ-mediated repair. Nevertheless, this system is limited to individual tests. Here we aim to generate a novel lentiviral dual-gRNA expression system that enables large scale functional enhancer screening. The candidate will apply this dual-gRNA expression system to generate custom libraries that will allow the functional identification and subsequent validation of essential enhancers for the maintenance of mouse Embryonic Stem (mES) cell identity. This technology will provide a comprehensive view of the function of enhancers within their genomic context and will enable for the first time the performance of systematic, genetic forward enhancer screens.

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Keywords

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Programme(s)

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Topic(s)

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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.

MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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

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Coordinator

THE UNIVERSITY OF EDINBURGH
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
OLD COLLEGE, SOUTH BRIDGE
EH8 9YL Edinburgh
United Kingdom

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Region
Scotland Eastern Scotland Edinburgh
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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