CRISPR-Associated Transposase Systems for Precise Genome Insertion
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Solution Overview
Problem
Current genome editing technologies lack affordable, easy-to-set-up, and scalable strategies for precise targeting of multiple positions within the eukaryotic genome, limiting their application in genome engineering and biotechnology.
Innovation Solution
An engineered nucleic acid targeting system comprising CRISPR-associated transposase proteins, Cas proteins, and guide molecules for sequence-specific binding and insertion of donor polynucleotides into target sequences, utilizing components like TnsB, TnsC, TniQ, and Cas12k to direct precise genome modifications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional genome editing technologies (designer zinc fingers, TALEs, homing meganucleases) are used, then targeted genome perturbations can be achieved, but the methods are complex, expensive, and difficult to scale for multiple positions
Solution Approach 1:
The patent combines CRISPR-Cas systems with transposase enzymes to create a hybrid system that merges the sequence-specific targeting capability of guide RNA-Cas complexes with the DNA insertion capability of transposases. This integration allows the system to achieve precise targeting while using a simpler, more scalable molecular mechanism than conventional genome editing technologies.
Solution Approach 2:
The patent uses guide RNA as an intermediary molecule that mediates between the Cas protein and the target DNA sequence. The guide RNA provides sequence-specific recognition and directs the Cas-transposase complex to precise genomic locations, enabling accurate targeting without requiring complex protein-DNA recognition systems like zinc fingers or TALEs.
2Measurement precision
If conventional genome editing technologies are used, then targeted genome perturbations can be achieved, but the methods are not affordable and easy to set up
Solution Approach 1:
The patent segments the genome editing function into separate modular components: the Cas protein provides targeting capability, the guide RNA provides sequence specificity, and the transposase provides insertion function. This modular segmentation allows each component to be independently optimized and assembled, making the system easier to set up and more affordable than integrated conventional methods.
Solution Approach 2:
The patent creates a universal platform where the Cas-transposase complex can target multiple genomic positions by simply changing the guide RNA sequence. This multi-functionality allows the same core system to be applied across numerous different target sites, reducing setup complexity and cost compared to designing separate systems for each target.
3Measurement precision
If conventional genome editing technologies are used, then targeted genome perturbations can be achieved, but scalability for multiple positions is limited
Solution Approach 1:
The patent enables scalable targeting of multiple positions by changing the parameter of guide RNA sequence rather than redesigning the entire editing system. Each guide RNA can be designed to target a specific genomic location, allowing rapid adaptation to new targets and efficient scaling to multiple positions while maintaining precise targeting capability.
Data Source
AI summary
The present application provides systems, methods and compositions used for targeted gene modification, targeted insertion, perturbation of gene transcripts, nucleic acid editing. Novel nucleic acid targeting systems comprise components of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems and transposable elements.