Dendronised Polymer Non-Viral Transfection Agent
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Solution Overview
Problem
Current gene delivery systems for genome editing tools, such as CRISPR/Cas9, face challenges including limited packaging capacity, toxicity, and low transfection efficiency, particularly for large plasmids, due to the limitations of viral and non-viral vectors like liposomes and dendrimers.
Innovation Solution
Development of biocompatible dendronised polymers with a linear aliphatic copolymer backbone and pendant PAMAM dendrons, formed through click chemistry, which combine the multi-valency of dendrimers with the conformational flexibility of linear polymers for optimal biomolecule binding and delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If dendrimer generation is increased to improve transfection efficiency, then the ability to deliver large genome editing DNA constructs is improved, but cytotoxicity increases significantly
Solution Approach 1:
The invention segments the dendrimer structure by attaching individual dendron units to a linear polymer backbone, creating a dendronised polymer. This segmentation allows the polymer to exhibit dendrimer-like transfection efficiency while maintaining the flexibility and lower toxicity of linear polymers, thus resolving the contradiction between transfection efficiency and cytotoxicity
Solution Approach 2:
The invention creates a composite material by combining linear polymer chains with dendron units. The resulting dendronised polymer integrates the advantages of both linear polymers (flexibility, low toxicity) and dendrimers (high transfection efficiency, multivalency), effectively resolving the technical contradiction
2Reliability
If dendrimer generation is increased to improve binding capacity, then the ability to form stable polyplexes with pDNA is improved, but conformational flexibility is reduced
Solution Approach 1:
By segmenting the rigid dendrimer structure into separate dendron units attached to a flexible linear backbone, the invention maintains polyplex stability through dendron-pDNA interactions while preserving the conformational flexibility of the linear polymer chain, thus resolving the contradiction between polyplex stability and conformational flexibility
3Reliability
If viral vectors are used to improve delivery capacity, then the ability to deliver multiple components is improved, but packaging capacity is limited
Solution Approach 1:
The dendronised polymer serves multiple functions: it can deliver various genome editing tools (CRISPR/Cas9, TALEs, zinc finger proteins), different types of nucleic acids (plasmids, siRNA), and can be used for both in vitro and in vivo applications. This multi-functionality allows it to replace multiple specialized viral vectors, effectively resolving the contradiction between delivery capacity and packaging capacity
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 dendronised polymers achieve high transfection efficiency and reduced cytotoxicity, enabling the effective intracellular delivery of large genome editing tools, comparable to commercial agents like Lipofectamine, while overcoming the limitations of traditional dendrimers.
Implementation Method 1
PAMAM dendrimers can act as efficient delivery agents due to a high density of primary amines on their periphery, which can interact with anionic DNA molecules to form stable polyplexes
Implementation Method 2
a high density of tertiary amines in their interior, which provide sufficient buffering capacity to enable endosomal escape of delivered DNA
Data Source
AI summary
The invention relates in general to hydrophilic, biocompatible dendronised polymers, to complexes comprising the dendronised polymers, to methods of preparing the dendronised polymer and to uses of the dendronised polymer as a non-viral transfection agent for the delivery of biomolecules, in particular, genome editing tools, into a cell.