Projektbeschreibung
Neuer Ansatz zur Untersuchung der allosterischen Regulierung im CRISPR-Cas9-Nukleoproteinkomplex
Die allosterische Regulierung steht in der Biochemie für die Regulierung eines Enzyms, indem ein Effektormolekül an einer anderen Stelle als der aktiven Stelle des Enzyms gebunden wird. Effektoren, welche die Proteinaktivität erhöhen, werden als „allosterische Aktivatoren“ bezeichnet und sind das Gegenteil von „allosterischen Inhibitoren“, welche die Proteinaktivität verringern. CRISPR-Cas9 ist ein großer Nukleoproteinkomplex, der gemeinhin als Genomeditierungswerkzeug verwendet wird. Sein beeindruckender allosterischer Signalweg beinhaltet das Multidomäne-Protein Cas9 und die damit verbundenen Nukleinsäuren, welche die Funktion und Spezifizität des Systems steuern. Das EU-finanzierte Projekt Allosteric-CRISPR erforscht die allosterische Regulierung bei CRISPR-Cas9, indem ein neuartiger synergistischer Ansatz als aufstrebendes Werkzeug für die Allosterieuntersuchung eingeführt wird. Dieser Ansatz kombiniert modernste theoretische Methoden mit Netzwerkmodellen, die von der Graphentheorie abgeleitet sind.
Ziel
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.
Wissenschaftliches Gebiet
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesmathematicspure mathematicsdiscrete mathematicsgraph theory
- natural sciencesbiological sciencesgeneticsgenomes
Schlüsselbegriffe
Programm/Programme
Thema/Themen
Finanzierungsplan
ERC-STG - Starting GrantGastgebende Einrichtung
80333 Muenchen
Deutschland