Descrizione del progetto
Un nuovo approccio per studiare la regolazione allosterica nel complesso nucleoproteico CRISPR-Cas9
In biochimica, la regolazione allosterica è la regolazione di un enzima effettuata legando una molecola, detta effettore, a un sito diverso dal sito attivo dell’enzima. Gli effettori che migliorano l’attività della proteina sono noti come «attivatori allosterici», in contrasto con gli «inibitori allosterici» che diminuiscono l’attività della proteina. CRISPR-Cas9 è un grosso complesso nucleoproteico, ampiamente usato come strumento di modifica del genoma. La sua intricata segnalazione allosterica coinvolge la proteina multidominio Cas9 e gli acidi nucleici associati, che controllano il funzionamento e la specificità del sistema. Il progetto Allosteric-CRISPR, finanziato dall’UE, indagherà sulla regolazione allosterica in CRISPR-Cas9 introducendo un approccio sinergico innovativo come potente strumento emergente per studiare l’allosteria. Tale approccio combina metodi teorici all’avanguardia con modelli di rete derivati dalla teoria dei grafi.
Obiettivo
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.
Campo scientifico
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesmathematicspure mathematicsdiscrete mathematicsgraph theory
- natural sciencesbiological sciencesgeneticsgenomes
Parole chiave
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-STG - Starting GrantIstituzione ospitante
80333 Muenchen
Germania