Cas Protein Epitope Mutations for Reduced Immunogenicity
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Solution Overview
Problem
Current genome-editing technologies, such as CRISPR-Cas systems, face challenges in reducing immunogenicity, which hinders their translation into clinical applications due to immune responses triggered by Cas proteins.
Innovation Solution
Mutating immunogenic T cell epitopes of Cas proteins using methods like phage display and phage-assisted continuous evolution (PACE) to reduce their immunogenicity while maintaining nuclease activity and target specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Cas proteins are used for genome editing, then genome editing capability is improved, but immunogenicity increases causing immune responses
Solution Approach 1:
The patent extracts and removes immunogenic epitopes from Cas proteins through site-directed mutagenesis. Specific amino acid sequences that trigger immune responses are identified and replaced with non-immunogenic variants, thereby separating the harmful immunogenic properties from the beneficial genome-editing function of the Cas protein
Solution Approach 2:
The patent applies local quality modification by making specific point mutations at immunogenic epitope locations while leaving the rest of the Cas protein structure unchanged. This targeted approach modifies only the problematic local regions (epitopes) to reduce immunogenicity while preserving the overall functionality and structure of the Cas protein
2Object-affected harmful factors
If T cell epitopes are mutated to reduce immunogenicity, then immunogenicity is reduced, but nuclease activity may be affected
Solution Approach 1:
The patent applies partial action by mutating only specific immunogenic epitopes rather than making comprehensive changes to the entire Cas protein. This selective mutation approach reduces immunogenicity through targeted modifications while minimizing disruptions to the overall protein structure and nuclease activity
Solution Approach 2:
The patent changes specific amino acid parameters at epitope locations through point mutations. By modifying individual amino acid residues that constitute immunogenic epitopes, the patent alters the immunogenic parameters of the protein while maintaining the structural and functional parameters necessary for nuclease activity
3Object-affected harmful factors
If multiple epitopes are mutated, then immunogenicity is reduced, but protein structure may be compromised
Solution Approach 1:
The patent segments the Cas protein into distinct functional regions and immunogenic epitopes. By identifying and separately mutating specific epitopic segments rather than making global changes, the patent reduces immunogenicity through localized modifications while preserving the integrity of the overall protein structure and its essential functional domains
Data Source
AI summary
The present disclosure generally relates to systems, methods and compositions related to Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and components thereof. The present disclosure also relates to methods, systems, and compostions modified to reduce immunogenicity. Additionally, the present disclosure relates to methods for developing or designing CRISPR-Cas system based therapy or therapeutics.