Project description
Novel approach to study allosteric regulation in CRISPR-Cas9 nucleoprotein complex
Allosteric regulation in biochemistry is the regulation of an enzyme by binding an effector molecule at a site other than the active site of the enzyme. Effectors that enhance the protein's activity are called 'allosteric activators', in contrast to 'allosteric inhibitors' that decrease the protein's activity. CRISPR-Cas9 is a large nucleoprotein complex, widely employed as a genome editing tool. Its intricate allosteric signaling involves the multi-domain Cas9 protein and its associated nucleic acids, controlling the system's function and specificity. The EU-funded Allosteric-CRISPR project will investigate the allosteric regulation in CRISPR-Cas9 by introducing a novel synergistic approach as an emerging powerful tool to investigate allostery. This approach combines state-of-the-art theoretical methods with network models derived from graph theory.
Objective
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.
Fields of science
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesmathematicspure mathematicsdiscrete mathematicsgraph theory
- natural sciencesbiological sciencesgeneticsgenomes
Programme(s)
Topic(s)
Funding Scheme
ERC-STG - Starting GrantHost institution
80333 Muenchen
Germany