CRISPR-Cas Nanoparticle Delivery for Genome Editing
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Solution Overview
Problem
Current genome editing technologies, such as designer zinc fingers and TALEs, are complex, costly, and not easily scalable for targeting multiple positions within the eukaryotic genome, necessitating a more affordable and efficient method for precise genome engineering.
Innovation Solution
The CRISPR-Cas system, which uses a single Cas enzyme programmed by a short RNA molecule to recognize specific DNA targets, is employed for genome editing, facilitated by nanoparticle delivery of a CRISPR complex comprising a guide sequence hybridized to a target sequence, enabling modifications like deletion, insertion, or activation of polynucleotides in various cell types and tissues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If traditional genome editing technologies (designer zinc fingers, TALEs) are used, then precise genome targeting is achieved, but device complexity and cost increase significantly
Solution Approach 1:
The CRISPR system divides the genome targeting function into two separate components: a Cas enzyme that performs the cutting action and a guide RNA that provides sequence specificity. This segmentation allows each component to be optimized independently and simplifies the overall system compared to traditional methods that require customized protein complexes for each target sequence
Solution Approach 2:
A single Cas enzyme can be programmed to target multiple different genomic sequences by simply changing the guide RNA sequence. This universal approach eliminates the need to develop different protein complexes for each target, reducing both device complexity and cost while maintaining precise genome targeting capability
2Manufacturing precision
If traditional genome editing technologies are used, then genome perturbation is achieved, but ease of manufacture and scalability deteriorate
Solution Approach 1:
The guide RNA can be easily synthesized and copied for multiple targets using standard molecular biology techniques. Unlike traditional methods that require de novo protein engineering for each target, the CRISPR system allows rapid replication of the same Cas enzyme with different guide RNAs, significantly improving ease of manufacture and scalability
Solution Approach 2:
The system allows easy modification of the guide RNA sequence to change target specificity without altering the Cas enzyme structure. This parameter change approach enables rapid adaptation to different genomic targets, making the system highly scalable and easy to manufacture for multiple applications
3Manufacturing precision
If CRISPR-Cas system is delivered without optimized delivery methods, then genome editing components are provided, but delivery efficiency and cell-type specific targeting are insufficient
Solution Approach 1:
The patent introduces particle delivery components as intermediaries to transport the CRISPR-Cas system into target cells. These particles serve as mediators that protect the genetic components during delivery and facilitate their entry into specific cell types, thereby improving delivery efficiency while maintaining the genome editing function
Solution Approach 2:
The delivery system is optimized for cell-type and tissue-specific targeting, providing local quality enhancement. By tailoring the particle delivery components to specific target cells, the system achieves high delivery efficiency in the intended tissue while minimizing off-target effects, thus resolving the contradiction between maintaining editing function and improving delivery productivity
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
This approach simplifies genome editing, reduces costs, and enhances scalability by allowing targeted modifications across multiple genomic locations, with applications in gene editing, therapy, drug discovery, and disease diagnosis, while minimizing off-target effects through precise sequence specificity.
Implementation Method 1
a guide sequence hybridized to a target sequence within the target polynucleotide
Data Source
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AI summary
The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery particle formulations and/or systems comprising one or more components of a CRISPR-Cas system, which are means for targeting sites for delivery. The delivery particle formulations of the invention are preferably nanoparticle delivery formulations and/or systems. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.