Compact Type V-G CRISPR Effector for Versatile Genome Editing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current CRISPR-Cas systems have limitations in modifying nucleic acids and polynucleotides, and there is a need for additional programmable effectors and systems that can enable novel applications through their unique properties.
Innovation Solution
Development of new single-effector Class 2 CRISPR-Cas systems, including the CLUST.019143 (Type V-G) CRISPR-Cas effector protein, which features a unique domain organization and provides novel DNA/RNA editing properties, smaller size for versatile delivery strategies, genotype-triggered cellular processes, and programmable RNA-guided DNA insertion, excision, and mobilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing CRISPR-Cas systems are used for genome editing, then DNA modification capability is achieved, but the system size is large and limits delivery versatility
Solution Approach 1:
The patent segments the CRISPR system into minimal essential components: a compact Type V-G effector protein (smaller than traditional Cas9), a simplified guide RNA structure, and separates the DNA-binding function from the cleavage function. This segmentation enables smaller protein size for improved delivery while maintaining editing capability.
Solution Approach 2:
The patent extracts and removes non-essential domains from traditional CRISPR-Cas systems, retaining only the core functional elements required for targeting and cleavage. This extraction reduces the overall system size and complexity, facilitating versatile delivery across different platforms.
2Adaptability or versatility
If multiple effector proteins are used in Class 1 CRISPR systems, then complex DNA modification capabilities are achieved, but the system complexity increases
Solution Approach 1:
The patent merges multiple effector protein functions into a single Type V-G effector protein that performs both DNA binding (via guide RNA complex formation) and DNA cleavage (via RuvC domain). This consolidation reduces component complexity from multiple separate proteins to a single integrated effector.
Solution Approach 2:
The Type V-G effector protein serves multiple functions: it complexes with guide RNA for target recognition, performs site-specific DNA cleavage through its RuvC domain, and can be programmed to target different sequences by changing the guide RNA. This multi-functionality replaces the need for multiple specialized effector proteins.
3Reliability
If traditional CRISPR systems are engineered for specific applications, then application-specific optimization is achieved, but the time and resources for engineering increase
Solution Approach 1:
The patent performs preliminary engineering by optimizing the Type V-G effector protein and guide RNA complex formation in advance, establishing a robust baseline system that requires minimal further engineering for specific applications. The pre-optimized system reduces the time and resources needed for subsequent application-specific adaptations.
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 new CRISPR-Cas systems enable broad applications for specific, programmed perturbations in genome and epigenome manipulation, offering enhanced editing capabilities and versatility.
Implementation Method 1
a CLUST.019143 (Type V-G) CRISPR-Cas effector protein... capable of binding to the RNA guide and of targeting the target nucleic acid sequence complementary to the spacer sequence
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 nucleic acids such as DNA. Each system includes one or more protein components and one or more nucleic acid components that together target nucleic acids.