Engineered Type VI-D CRISPR-Cas Effector Proteins
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Solution Overview
Problem
Current CRISPR-Cas systems have limitations in providing additional programmable effectors for nucleic acid modification beyond existing systems, necessitating the development of new Class 2 CRISPR systems with enhanced capabilities for genome engineering and biotechnological applications.
Innovation Solution
The development of engineered CRISPR-Cas systems, including RNA guides and Type VI-D CRISPR-Cas effector proteins with specific amino acid sequences and accessory proteins, capable of binding to target nucleic acids and modulating their activity, enabling targeted nucleic acid modification and cleavage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If new Class 2 CRISPR systems are developed to expand programmable effector capabilities, then the versatility and scope of genome engineering applications are improved, but the device complexity and difficulty of system engineering increase
Solution Approach 1:
The patent divides the CRISPR system into distinct functional modules: RNA guide components (crRNA with direct repeat and spacer sequences), effector proteins (Cas13d variants with HEPN domains), and accessory proteins (WYL domain proteins). This segmentation allows independent optimization and engineering of each component while maintaining overall system functionality, thereby expanding versatility without proportionally increasing complexity.
Solution Approach 2:
The patent develops universal CRISPR system architecture where the effector protein can work with different RNA guide sequences to target various nucleic acid sequences. The standardized interface between RNA guides and effector proteins allows a single effector protein to perform multiple targeting functions, enhancing versatility while controlling complexity through reusability.
2Manufacturing precision
If engineered CRISPR systems with specific amino acid sequences are designed to enhance targeting efficiency, then the manufacturing precision and specificity of nucleic acid modification are improved, but the ease of manufacture and protein engineering difficulty increase
Solution Approach 1:
The patent introduces specific amino acid substitutions at localized positions within the effector protein (e.g., HEPN domain residues) to enhance catalytic activity and targeting specificity. Rather than redesigning the entire protein, localized mutations are made to specific functional regions, improving precision while maintaining the overall protein structure and simplifying the engineering process.
Solution Approach 2:
The patent systematically varies amino acid parameters (charge, hydrophobicity, size) at key positions to optimize protein-RNA interactions and catalytic efficiency. By changing specific physical-chemical parameters of amino acids at critical positions, the patent achieves enhanced targeting precision while using standard protein engineering approaches that remain relatively straightforward to implement.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
These engineered systems provide enhanced programmability and efficiency in nucleic acid targeting and modification, expanding the scope of genome engineering applications and biotechnological uses.
Implementation Method 1
a spacer sequence capable of hybridizing to a target nucleic acid
Implementation Method 2
the effector protein is capable of binding to the RNA guide and of targeting the target nucleic acid
Implementation Method 3
engineered systems for modifying nucleic acids and polynucleotides
Data Source
AI summary
The disclosure describes novel systems, methods, and compositions for the manipulation of nucleic acids in a targeted fashion. The disclosure describes non-naturally occurring, engineered CRISPR systems, components, and methods for targeted modification of a nucleic acid.