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Identifying Variable Chromatin Modules using single-cell epigenomics

Project description

A method for studying the interplay of distant genomic regions

Gene expression is regulated through the complex interplay between cis-regulatory sequences and trans-acting elements located at distant sites. Dynamic changes in the activity of these elements underlie phenotypic variation and disease. To explain the molecular coordination between genomic regions spanning over 100 kb researchers have come up with the concept of variable chromatin modules (VCMs). The EU-funded CHROMISE project aims to determine VCM function and plasticity through a novel experimental and computational workflow that makes VCM identification fast and easy. Researchers will use the differentiation of mesenchymal stem cells as a model system and apply the generated knowledge to study genetic variants of metabolic diseases.

Objective

Most common disease-associated genetic variants are thought to fall into gene regulatory regions, where they affect the interaction between transcription factors (TFs) and DNA and induce transcriptional changes. To understand the extent of, and the mechanisms underlying variation in binding of TFs to DNA, genome-wide TF and chromatin state profiling studies were performed which revealed that non-coding variants frequently mediate coordinated changes in TF binding and chromatin mark enrichment over regions that span more than 100 kb. These regions were termed “variable chromatin modules” (VCMs), providing a conceptual framework of how regulatory variation might shape complex traits. However, little is known about the formation, function, or plasticity of VCMs. This is because vast amounts of epigenomics data have so far been required for identifying VCMs, making their identification costly, labour-intensive, and only applicable to easy-to-culture cell types. Recent work has demonstrated the value of ATAC-seq for defining regulatory element hierarchies that conceptually resemble VCMs. Here, I propose to extend these principles by developing an experimental and computational workflow for VCM identification using single-cell ATAC-seq. This will allow sample multiplexing for sequencing followed by genotype-based demultiplexing, thereby increasing throughput, minimizing sample input, and mitigating variation between samples (Aim 1). I will apply this workflow to study VCM plasticity during human mesenchymal stromal cell differentiation (Aim 2). The resulting data will be used to identify VCM-linked metabolic disease-relevant genetic variants, whose role in VCM formation will be validated using CRISPR-Cas9 (Aim 3).

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Topic(s)

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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-2020

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Coordinator

ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE
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.

€ 203 149,44
Address
BATIMENT CE 3316 STATION 1
1015 LAUSANNE
Switzerland

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Region
Schweiz/Suisse/Svizzera Région lémanique Vaud
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.

€ 203 149,44
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