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Large scale perturbations of the gene regulatory networks of E. coli

Project description

Complex study of the gene regulatory network in bacteria

The EU-funded PERTURBATIONS project will study the relationships between genotype, phenotype, fitness and environment in a model bacterial system using a microfluidic technology coupled with transcriptomics, aiming to decipher gene regulatory networks, evolution, and evolutionary constraints. The research will involve programmable DNA altering CRISPR technology, suitable to perturb gene expression of combinatorial global regulators of the transcription, cascades of genes in the network hierarchy, and single global regulators with mutations at many positions. The expression patterns of the transcriptome will be quantified using RNA sequencing. The fitness of each strain will be quantified using microscopy and image analysis. The cutting-edge microfluidic system will enable the study of many strains of bacteria in a high-throughput manner in a strictly controlled environment.

Objective

Although genome sequencing is well advanced, our understanding of the genome to phenotype to fitness relationship remains very poor. It is becoming very evident that the information stored in genes is not enough to understand their impact without knowing the phenotypic and environmental context. The fact that a gene is present in a genome tells very little about when it is expressed and its relationship to other genes and mutual effects on gene expression pattern. To understand how phenotypes emerge from a specific genotype, we need to understand gene regulatory networks and their mechanisms of regulation. In this project, I propose to study the relationships between genotype, phenotype, fitness, and environment using a microfluidic device coupled with transcriptomics with the goal of collecting high dimension high quality data for unprecedented level of understanding and predicting regulatory networks, evolution, and evolutionary constraints in the form of epistasis and pleiotropy. I will use programmable DNA binding protein called CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) to perturb gene expression of (1) combinatorial global regulators within the transcriptional regulatory network, (2) cascades of genes that form gene network hierarchy, and (3) single global regulators with low level mutations at many positions in their genetic code. I will then quantify the expression pattern of the transcriptome using RNA sequencing and the growth fitness of each strain. The environment will be highly controllable as we will create a cutting-edge microfluidic system to study many strains of bacteria in a high-throughput manner. Successful implementation of the strategies explored in this project will open the doors to plenty of other avenues to explore with fruitful outcomes wherever there is a need for combining genotypic, phenotypic, and fitness studies in highly controllable environments

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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 SUPERIEURE DE PHYSIQUE ET DECHIMIE INDUSTRIELLES DE LA VILLE DEPARIS
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.

€ 184 707,84
Address
RUE VAUQUELIN 10
75231 Paris
France

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Region
Ile-de-France Ile-de-France Paris
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.

€ 184 707,84
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