Description du projet
Une nouvelle approche pour étudier la régulation allostérique du complexe nucléoprotéique CRISPR/Cas9
La régulation allostérique en biochimie concerne la régulation d’une enzyme par la liaison d’une molécule effectrice à un site autre que le site actif de l’enzyme. Les effectrices qui renforcent l’activité protéique sont appelées «activatrices allostériques», contrairement aux «inhibitrices allostériques» qui diminuent l’activité de la protéine. CRISPR/Cas9 désigne un grand complexe nucléoprotéique, largement employé en tant qu’outil d’édition génomique. Sa signalisation allostérique complexe implique la protéine multi‑domaine Cas9 ainsi que ses acides nucléiques associés, contrôlant la fonction et la spécificité de ce système. Le projet Allosteric‑CRISPR, financé par l’UE, étudiera la régulation allostérique du CRISPR/Cas9 en introduisant une nouvelle approche synergique en tant que puissant outil émergent pour étudier l’allostérie. Cette approche combine des méthodes théoriques de pointe avec des modèles de réseaux dérivés de la théorie des graphes.
Objectif
Allostery is a fundamental property of proteins, which regulates biochemical information transfer between spatially distant sites. Many emerging allosteric targets are large protein/nucleic acid complexes responsible for genome editing and regulation, whose underlying signaling remains poorly understood. Here, we focus on CRISPR-Cas9, a large nucleoprotein complex widely employed as a genome editing tool with enormous promises for medicine and biotechnology. In this system, an intricate allosteric signaling is suggested to span the multi-domain Cas9 protein and its associated nucleic acids, controlling the system’s function and specificity. However, in spite of extensive experimental characterization, the molecular basis for this allosteric response are largely unknown, hampering also efficient engineering for improving its genome editing capability. Allosteric-CRISPR will investigate the allosteric regulation in CRISPR-Cas9 by introducing a novel synergistic approach. This will implement the combination of state-of-the-art theoretical methods, including enhanced and multiscale approaches based on classical and ab-initio methods, with network models derived from graph theory and novel centrality analyses that are emerging as powerful to investigate allostery. This will create an innovative protocol that will enable determining the allosteric network of communication over multiple timescales, as well as the relation between allostery and catalysis, which remains unaddressed through classical approaches. This novel way to describe allostery can impact future studies of large nucleoprotein complexes, including newly discovered CRISPR systems, which are governed by similar allosteric rules and hold tremendous potential for genome editing. Finally, by delivering fundamental knowledge on the basic mechanisms underlying genome editing, Allosteric-CRISPR will help the design of improved genome editing tools, impacting their application across the field of life sciences.
Champ scientifique
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesmathematicspure mathematicsdiscrete mathematicsgraph theory
- natural sciencesbiological sciencesgeneticsgenomes
Programme(s)
Thème(s)
Régime de financement
ERC-STG - Starting GrantInstitution d’accueil
80333 Muenchen
Allemagne