Hyperbranched Polymer Nanoparticles for CRISPR Delivery
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Solution Overview
Problem
Current methods for delivering CRISPR genomic editing for treating skin genetic disorders like RDEB face challenges in safety and efficacy due to immune responses and off-target effects, requiring a more efficient and cytocompatible delivery system.
Innovation Solution
The use of hyperbranched poly(beta-amino ester) polymers to condense ribonucleoprotein complexes into nanoparticles that can efficiently penetrate cells and correct type VII collagen mutations in RDEB, achieving high transfection and correction efficiency with minimal cytotoxicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral vectors are used to deliver CRISPR into cells, then delivery efficiency is improved, but immune response and off-target effects increase
Solution Approach 1:
The patent uses a non-viral polymer delivery vector as an intermediary to transport CRISPR components into cells. This polymer-based mediator avoids the immune recognition and off-target effects associated with viral vectors while maintaining delivery capability, thus resolving the contradiction between delivery efficiency and immune response
Solution Approach 2:
The patent employs transient expression of therapeutic Cas9 and guide RNAs via non-viral delivery, using short-lived polymer complexes instead of persistent viral vectors. This approach achieves the necessary delivery effect without the long-term safety concerns of viral integration, reducing both immune response and off-target effects
2Object-affected harmful factors
If non-viral polymer delivery vectors are used, then immune response is reduced, but delivery efficiency decreases
Solution Approach 1:
The patent modifies the parameters of non-viral polymer vectors by using hyperbranched poly(beta-amino ester) polymers with specific cationic charges and molecular structures. These parameter changes enhance the transfection efficiency and cellular uptake of the CRISPR components, overcoming the traditional efficiency limitations of non-viral delivery while maintaining low immunogenicity
3Ease of operation
If CRISPR-plasmid systems are used, then delivery is achieved, but transfection efficiency and correction efficiency are lower
Solution Approach 1:
The patent changes the delivery vehicle from plasmid-based to ribonucleoprotein (RNP) complex-based CRISPR delivery. By complexing Cas9 protein with guide RNA and delivering as RNP particles, the system achieves significantly higher transfection efficiency and correction efficiency (8.2% to 43.2%) compared to plasmid systems, while maintaining delivery capability
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
The nanoparticulate compositions demonstrate higher transfection and correction efficiency compared to traditional CRISPR-plasmid systems, effectively excising exon 80 and restoring type VII collagen in RDEB patients with improved cell viability and reduced side effects.
Implementation Method 1
hyperbranched poly (beta-amino) ester polymers are capable of efficiently condensing ribonucleoprotein complexes into nanoparticles
Implementation Method 2
facilitate transport across the cell membrane in an efficient and cytocompatible manner
Data Source
AI summary
A nanoparticulate composition comprises a gene editing ribonucleoprotein system complexed within a cationic polymer. The cationic polymer may be a Poly-beta amino ester hyperbranched polymer, especially a 4-branching hyperbranched polymer. The gene editing ribonucleoprotein system may be a CRISPR-Cas9 gene editing system configured to excise a mutation or exon in a gene, replace a mutation in a gene, or produce a knock-down or knock-out of a gene, and in particular configured to excise exon 80 of the COL7A1 gene which codes for the collagen VII protein. Data shows that using