Polysaccharide Nanoparticles for CRISPR-Cas Delivery Stability
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Solution Overview
Problem
Current CRISPR-Cas-based technologies face challenges in delivering the CRISPR-Cas-associated complex effectively into human cells, as the complex is unstable in the bloodstream and lacks a suitable delivery vehicle, leading to inefficiencies and potential side effects.
Innovation Solution
The development of a nanoencapsulation method using a biocompatible polysaccharide shell to encase the CRISPR-Cas system, specifically using ionic polysaccharides like chitosan and trimethyl chitosan with tripolyphosphate, forming stable nanoparticles that protect the CRISPR-Cas complex and facilitate cellular uptake.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CRISPR-Cas system is delivered directly into human cells, then the antiviral therapy can be effective, but the complex is unstable in the bloodstream and lacks suitable delivery vehicle
Solution Approach 1:
The patent uses polysaccharide-based nanoparticles as an intermediary delivery vehicle to transport the CRISPR-Cas complex into human cells. The nanoparticles serve as a mediator between the unstable CRISPR-Cas complex and the target cells, providing both stability during bloodstream circulation and efficient cellular uptake. This resolves the contradiction by introducing a stable carrier that protects the complex while enabling effective delivery.
Solution Approach 2:
The CRISPR-Cas complex is encapsulated within the polysaccharide nanoparticle structure, creating a nested configuration where the complex is protected inside the nanoparticle shell. This nesting approach provides stability during circulation while maintaining the ability to deliver the complex into target cells, effectively resolving the stability-delivery contradiction.
2Productivity
If CRISPR-Cas complex is used for antiviral therapy, then specific viral RNA sequences can be targeted, but the complex degrades in the bloodstream
Solution Approach 1:
The patent employs a polysaccharide shell (flexible membrane) to encapsulate the CRISPR-Cas complex. This shell provides protection against degradation by nucleases in the bloodstream while maintaining flexibility for cellular uptake. The shell acts as a protective barrier that preserves the complex's integrity during circulation, directly addressing the stability issue without compromising antiviral efficiency.
Solution Approach 2:
The invention uses composite polysaccharide materials (such as chitosan, alginate, or hyaluronic acid) to create nanoparticles that combine structural stability with biocompatibility. These composite materials form a protective matrix that shields the CRISPR-Cas complex from degradation while enabling efficient cellular internalization, thus resolving the contradiction between stability and productivity.
3Ease of operation
If conventional delivery methods are used for CRISPR-Cas, then the system can be administered, but cellular uptake is inefficient and side effects occur
Solution Approach 1:
The patent modifies key parameters of the delivery system by adjusting the size, charge, and surface properties of the polysaccharide nanoparticles. By controlling these parameters, the nanoparticles achieve optimal cellular uptake efficiency while maintaining ease of administration. The parameter optimization resolves the contradiction by creating a delivery system that is both easy to administer and highly efficient at cellular internalization.
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 creates a stable, biocompatible delivery vehicle that protects the CRISPR-Cas system from degradation and ensures efficient cellular uptake, potentially providing a effective antiviral therapy for conditions like COVID-19 by targeting specific viral RNA sequences.
Implementation Method 1
in which payload molecule zipped in between the two oppositely charged polysaccharide macromolecules
Implementation Method 2
wrapped by several layers of oppositely charged polysaccharides, so the payload is located in the centre of the formed nanoparticle, and is isolated from external attacks by RNAse and ferments
Data Source
AI summary
Nanoparticles capable of delivering active pharmaceutical compounds (payload) into cells and methods of preparing same; wherein the payload is ionized and combined with a first polymer ion of an opposite charge resulting in the formation of an initial molecular assembly. The initial molecular assembly is then combined with a second polymer ion having an opposite charge and a different size than the first polymer ion, resulting in the formation of a secondary molecular assembly, wherein the payload becomes trapped between the first and the second polymer ions. The non-conjugated charged segments of the secondary molecular assembly are then repeatedly combined with additional polymer ions of alternating opposite charges to such segments charges and pre-determined size to extend and branch the molecular assembly resulting in the formation of a nanoparticle.