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Development of a universal fluorescence sybody-based biosensor for the in-vivo study of protein dynamics

Project description

Nanobody sensors for cell monitoring

Genetically encoded fluorescent biosensors are specially designed proteins produced by genetically engineered cells that fluoresce to report specific biological activity, such as chemical changes or protein behaviour. These sensors allow scientists to monitor what’s happening inside living cells in real time, discover disease mechanisms and test new drugs. However, designing such biosensors is complex, and current options can only track a limited number of cellular processes. With the support of the Marie Skłodowska-Curie Actions programme, the UniverSy project proposes to develop a new flexible biosensor system built from nanobodies, small antibodies that can recognise very specific proteins. These molecules can serve as universal biosensors for tracking more biological targets.

Objective

The development of genetically-encoded fluorescence sensors revolutionized the monitoring of biochemical processes in living cell by reporting a change in their fluorescence properties in response to a specific aspect of their environment. However, the design of such biosensors is not trivial and available tools only target a limited set of phenomena. Current limitations prevent the monitoring of crucial processes such as the activity of membrane associated receptors which are a rich target for drug development. Simple binders are much easier to develop than biosensors, for instance nanobodies can bind a plethora of targets with high affinity and specificity, sometimes for a particular conformation. The host research group recently developed a molecular design to convert nanobodies into fluorescent biosensors. To elicite a fluorescent signal in presence of the target, this NanoBlock sensors require the development of an intracellular blocking peptide that will be displaced upon binding to the target. This limiting step prevent the application of the NanoBlock design to a large number of targets.
This objective of this proposal is to develop an entirely new class of fluorescent biosensors by engineering the NanoBlock technology so that nanobodies can be converted into fluorescent biosensors without the need for a distinct blocking peptide creating a plug and play system. This UniverSy technology will open the possibilities to visualize many more cellular processes to gain fundamental insight on protein activity and the development of therapeutics targeting these processes. The development of the UniverSy sensor will consist in the generation of dual affinity nanobodies binding their original target and a specific peptide sequence; the demonstration of the versatility of the UniverSy framework to target various proteins in vitro and in living cell and the use of UniverSy in a fluorescence assay to quantify the activation of GPCR for drug screening applications.

Fields of science (EuroSciVoc)

Keywords

Programme(s)

Topic(s)

Funding Scheme

HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

KATHOLIEKE UNIVERSITEIT LEUVEN
Net EU contribution
€ 216 240,00
Address
OUDE MARKT 13
3000 Leuven
Belgium
Region
Vlaams Gewest Prov. Vlaams-Brabant Arr. Leuven
Activity type
Higher or Secondary Education Establishments
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Total cost
No data