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Location, location, location: the (re-)positioning of regulatory elements in the mammalian genome.

Project description

How (re-)positioning of regulatory elements affects gene activity

In mammalian genomes, enhancers (Es) can be positioned nearby or far from the promoter (P) that they regulate. The EU-funded RE_LOCATE project aims to develop a technology to transplant selected DNA elements to hundreds of alternative positions, revealing how the ordering and spacing of Es and Ps contribute to gene regulation. The researchers will combine this technology with a high-throughput combinatorial reporter assay to study the role of Es selectivity in activation of the correct Ps. Also, the technology will be used to map the loop extrusion that shapes E and P interactions, and fine-map the repressive/activating chromatin landscape at high resolution. This work will reveal how the positioning of regulatory elements along the genome contributes to optimal gene regulation.

Objective

"In mammalian genomes, the enhancers (E) that control a promoter (P) are often scattered over tens to hundreds of kb, and frequently interdigitate with Es that control other nearby Ps. How such apparently haphazard linear arrangements can result in specific gene regulation is a major puzzle. Three main constraints are thought to be involved. First, biochemical compatibility of Es and Ps may ensure that not all Ps respond equally strongly to a given E. Second, chromatin loops may either facilitate or curb particular E-P interactions. Third, Es and Ps are controlled by the local landscape of chromatin modifications. Much is still to be learned about these constraints and their interplay. To unravel the logic of this linear arrangement of Ps and Es, it is necessary to systematically alter the positions of Es, Ps, and elements that control looping. So far, no efficient method has been available for this purpose. We propose to develop and apply RElocate, a scalable, broadly applicable technology to transplant selected DNA elements to hundreds of alternative positions within a ~2 Mb region, and track the functional consequences. We will employ RElocate in combination with a high-throughput combinatorial reporter assay to systematically study how biochemical compatibility may dictate how Es ""choose"" and activate the correct target P(s). Furthermore, we will adapt RElocate to precisely map how loop extrusion shapes E-P interactions, by insertion of hundreds of unidirectional road blocks throughout a locus. Finally, we will use the method to fine-map the repressive/activating chromatin landscape of selected regions at high resolution, and elucidate how Es and Ps may respond differently when inserted throughout this landscape. This work will reveal how the ordering and spacing of regulatory elements along the genome contributes to optimal gene regulation, and will yield a powerful perturbation tool with many applications in genome biology and human genetics."

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Programme(s)

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Topic(s)

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HORIZON-ERC - HORIZON ERC Grants

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Call for proposal

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(opens in new window) ERC-2021-ADG

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Host institution

STICHTING HET NEDERLANDS KANKER INSTITUUT-ANTONI VAN LEEUWENHOEK ZIEKENHUIS
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.

€ 2 493 875,00
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.

€ 2 493 875,00

Beneficiaries (1)

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