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Systematic identification of (p)ppGpp-dependent multidrug and stress tolerance factors

Objective

As a global healthcare issue, chronic and recurrent infections are often caused by multidrug tolerant persister cells, which are regulated by redundant pathways involving the almost ubiquitous alarmone (p)ppGpp. Despite extensive studies, two major questions in the persistence field remain to be answered, 1) the extent to which (p)ppGpp is involved in persistence and 2) how persistent cells resuscitate. To address these questions, three state-of-the-art techniques will be used in this project. First, DRaCALA (differential radial capillary action of ligand assay) will be used to systematically screen for proteins directly binding (p)ppGpp for the first time. The potential fellow from the Imperial College London has tremendous experience in DRaCALA. Second, barcode analysis by sequencing (Bar-seq) technique will be used to screen for persistence-specific and, for the first time, also resuscitation-specific genes. Collaboration with the partner Professor Duncan Maskel at the University of Cambridge, UK, a specialist in quantitative DNA deep sequencing, will ensure smooth progress of this objective and enable the potential fellow to be trained with this first-class technique. Both DRaCALA and Bar-seq will be transferred to the host lab. At last, identified candidate genes and proteins through the first two objectives will be validated with the contemporary single cell persistence assay developed in the host Professor Kenn Gerdes’s lab, at the University of Copenhagen, Denmark. Bar-seq and single cell persistence assay will be transferred to the potential fellow by senior postdocs in the partner and host labs, respectively. The tripartite cooperation with complementary expertise on a timely important project is expected to generate significant amount of high-quality data for the persistence field, facilitate extensive transfer of knowledge and promote the potential fellow to obtain an independent research position in the area of molecular microbiology and physiology.

Fields of science (EuroSciVoc)

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Topic(s)

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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-2015

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Coordinator

KOBENHAVNS UNIVERSITET
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.

€ 212 194,80
Address
NORREGADE 10
1165 KOBENHAVN
Denmark

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Region
Danmark Hovedstaden Byen København
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.

€ 212 194,80
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