Descripción del proyecto
Nuevo enfoque para el estudio de la regulación alostérica en el complejo nucleoproteico CRISPR-Cas9
En bioquímica, la regulación alostérica es la regulación de una enzima al enlazarse una molécula efectora en un sitio diferente al sitio activo de la enzima. Los efectores que mejoran la actividad de la proteína se denominan activadores alostéricos, a diferencia de los inhibidores alostéricos que disminuyen la actividad de la proteína. CRISPR-Cas9 es un gran complejo nucleoproteico, ampliamente utilizado como herramienta de edición genómica. Su señalización alostérica intrincada involucra a la proteína Cas9 de multidominio y sus ácidos nucleicos asociados, y controla la función y especificidad del sistema. El proyecto Allosteric-CRISPR, financiado con fondos europeos, investigará la regulación alostérica en el complejo CRISPR-Cas9 mediante la introducción de un enfoque sinérgico novedoso como una poderosa herramienta de nueva aparición para investigar la alostería. Este enfoque combina métodos teóricos pioneros con modelos de redes derivados de la teoría de grafos.
Objetivo
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.
Ámbito científico
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesmathematicspure mathematicsdiscrete mathematicsgraph theory
- natural sciencesbiological sciencesgeneticsgenomes
Palabras clave
Programa(s)
Régimen de financiación
ERC-STG - Starting GrantInstitución de acogida
80333 Muenchen
Alemania