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In Vitro Reconstitution of the Minimal Eukaryotic Cell Cycle Oscillator

Project description

In vitro biosensor platform for the cell cycle clock study

The experimental validation of the cell cycle clock/oscillations regulation by the cyclin-dependent kinase 1 represents a challenging task. The EU-funded CellCycleInVitro project proposes to answer questions about the cell cycle clock involving in vitro reconstitution as an innovative experimental approach. The combination of state-of-the-art biochemistry and synthetic and computational biology in application to the frog egg extract system will recreate the complete cell cycle oscillator in vitro and establish which elements of the oscillator define its frequency. This development of a novel in vitro biosensor platform for the study of cell division will provide new tools for applications ranging from synthetic biology to the development of new therapeutics.

Objective

Cell growth and division are the basis of all forms of life. In eukaryotes, the clock-like cycles of genome duplication and cell division are orchestrated by a complex set of biochemical reactions. Theory suggests that a negative feedback loop regulating the cyclin-dependent kinase 1 generates the cell cycle oscillations; however, because the putative cell cycle clock is tightly interlinked with numerous other biological networks, experimental validation has proven to be a challenge. To date, several questions key to understanding how cell cycle oscillations arise remain unanswered: Do the identified core components indeed give rise to oscillations? Which components are necessary and sufficient? And what makes the cell cycle clock oscillate with varying frequency?
To unravel what makes the cell cycle tick, we will reconstitute the core cell cycle oscillator in vitro. Specifically, combining state-of-the-art biochemistry, including the powerful frog egg extract system, with synthetic and computational biology, we will
(1) develop fluorescent-based sensors to monitor key enzymatic activities of the cell cycle oscillator,
(2) use computational models to identify operational conditions for the reconstitution of the oscillator,
(3) reconstitute and analyse the individual reactions of the predicted negative feedback loop, and
(4) assemble the complete cell cycle oscillator in vitro and establish its capacity to oscillate, the set of necessary components, and which elements of the oscillator define its frequency.
Using in vitro reconstitution as a radically new approach to interrogating the cell cycle, CellCycleInVitro will answer longstanding questions about the cell cycle clock. But more than this, in developing a novel in vitro experimental platform and biosensors for the study of cell division, we will provide powerful new tools with applications ranging from synthetic biology to the development of new therapeutics targeting uncontrolled cell proliferation.

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(opens in new window) ERC-2022-STG

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Host institution

RIJKSUNIVERSITEIT GRONINGEN
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.

€ 1 500 000,00
Address
Broerstraat 5
9712CP Groningen
Netherlands

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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.

€ 1 500 000,00

Beneficiaries (1)

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