pH-Responsive Nanoparticles for RNP Delivery
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Solution Overview
Problem
Current genome editing technologies, particularly those using CRISPR-Cas systems, face challenges in delivering ribonucleoproteins (RNPs) and single-stranded donor oligonucleotides (ssODNs) efficiently and safely due to their complex structures and susceptibility to degradation.
Innovation Solution
The development of self-assembled nanoparticles based on a pH-responsive amphiphilic polymer that accommodates RNPs and optionally ssODNs through electrostatic and hydrophobic interactions, providing high loading efficiencies and small uniform sizes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If RNP and ssODN are delivered without packaging, then delivery simplicity is improved, but stability and protection from degradation worsen
Solution Approach 1:
The patent introduces a pH-responsive amphiphilic polymer as an intermediary carrier that forms nanoparticles to encapsulate RNP and ssODN. This mediator protects the cargo from degradation while enabling cellular delivery, resolving the contradiction between simplicity and stability.
Solution Approach 2:
The polymer's pH-responsive properties allow it to change conformation and charge state in response to pH changes, enabling stable encapsulation in acidic environments while maintaining delivery functionality. This parameter change resolves the stability-simplicity contradiction.
2Reliability
If complex packaging systems are used to protect RNP and ssODN, then stability is improved, but device complexity worsens
Solution Approach 1:
The amphiphilic polymer self-assembles into nanoparticles through spontaneous aggregation driven by hydrophobic interactions and electrostatic forces. This self-service mechanism eliminates the need for complex external packaging systems while providing adequate protection.
Solution Approach 2:
The nanoparticle consists of a composite structure with a hydrophobic core containing RNP and ssODN, surrounded by a hydrophilic polymer shell. This composite material provides protection without requiring complex external packaging.
3Quantity of substance
If large nanoparticle sizes are used to accommodate RNP and ssODN, then loading capacity is improved, but cellular uptake efficiency worsens
Solution Approach 1:
The amphiphilic polymer forms a thin flexible shell around the RNP-ssODN core, providing adequate protection while maintaining a compact overall size that facilitates cellular uptake. The flexible shell adapts to the cargo size without significantly increasing the nanoparticle diameter.
4Productivity
If pH-responsive polymers are used to enable endosomal escape, then delivery efficiency is improved, but manufacturing precision requirements worsen
Solution Approach 1:
The polymer incorporates pH-responsive ionizable amine groups that automatically change charge state in response to pH changes in different cellular compartments. This inherent parameter change mechanism provides endosomal escape functionality without requiring precise manufacturing control.
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 nanoparticles offer a safe and efficient approach for both in vitro and in vivo genome editing, with good biocompatibility and low immunogenicity, enabling targeted gene editing with high precision.
Implementation Method 1
The RNP, ssODN, and the acrylic block of the amphiphilic copolymer form a core of the self-assembled nanoparticle
Implementation Method 2
accommodates the heterogeneity (e.g., charge and hydrophobicity) of RNP, and optionally ssODN, via both electrostatic and hydrophobic interactions
Implementation Method 3
the poly(ethylene glycol) block of the amphiphilic copolymer forms the exterior of the self-assembled nanoparticle
Implementation Method 4
an acrylic block comprising a poly(acrylate), poly(methacrylate) or poly(acrylate/methacrylate) block; the acrylic block comprise ester side chains bearing substituted or unsubstituted alkylamine groups
Data Source
AI summary
Provided herein are self-assembled nanoparticles (NPs), pharmaceutical compositions containing such NPs and methods of using such NPs. The NPs comprise an amphiphilic copolymer and a ribonucleoprotein (RNP), and optionally ssODN, wherein: the amphiphilic copolymer is a water-soluble block copolymer comprising a poly(C2-3 alkylene glycol) block and an acrylic block comprising a poly(acrylate), poly(methacrylate) or poly(acrylate/methacrylate) block; the acrylic block comprise ester side chains bearing substituted or unsubstituted alkylamine groups; and the RNP, ssODN, and the acrylic block of the amphiphilic copolymer form a core of the self-assembled nanoparticle, and the poly(ethylene glycol) block of the amphiphilic copolymer forms the exterior of the self-assembled nanoparticle.