CRISPR Nanocomplex Polymer Carrier Delivery
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Solution Overview
Problem
Current genome editing methods using CRISPR systems face challenges with low delivery efficiency and toxicity issues, particularly when delivering CRISPR enzyme proteins and sgRNA into bacterial cells, due to poor intracellular delivery and enzymatic degradation.
Innovation Solution
A CRISPR nanocomplex is formed by conjugating a polymer carrier material, such as branched polyethyleneimine, with the CRISPR enzyme protein and mixing it with sgRNA, enhancing delivery efficiency and minimizing toxicity by forming a stable, nonviral genome editing system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral vectors are used for CRISPR delivery, then transfection efficiency is improved, but cellular immune responses and cytotoxicity increase
Solution Approach 1:
The patent uses a polymer carrier (intermediary substance) to mediate the delivery of CRISPR components into cells, replacing viral vectors. The polymer carrier condenses sgRNA and Cas9 protein into nanocomplexes that can enter cells without triggering immune responses, thus resolving the contradiction between efficient delivery and immune safety
Solution Approach 2:
The patent changes the physical-chemical parameters of the delivery system by using polymers with specific properties (molecular weight, charge density, biocompatibility) to form nanocomplexes with controlled size and stability, achieving both efficient cellular uptake and reduced cytotoxicity compared to viral vectors
2Object-affected harmful factors
If nonviral carriers are used for CRISPR delivery, then cytotoxicity is reduced, but intracellular delivery efficiency decreases
Solution Approach 1:
The patent creates a composite nanocomplex system combining polymer carrier, sgRNA, and Cas9 protein. This composite structure leverages the advantages of each component: the polymer provides biocompatibility and low cytotoxicity, while the organized nanocomplex structure enables efficient cellular uptake and intracellular delivery, resolving the contradiction between safety and efficiency
3Device complexity
If CRISPR components are delivered without carrier protection, then system simplicity is maintained, but enzymatic degradation increases
Solution Approach 1:
The polymer carrier acts as a protective intermediary that shields the CRISPR components (sgRNA and Cas9 protein) from enzymatic degradation in the extracellular environment. The carrier maintains structural integrity during circulation and only releases the active components inside the cell, thus protecting against degradation while maintaining functional simplicity
Solution Approach 2:
The polymer carrier forms a protective shell around the CRISPR nanocomplex, providing a physical barrier against enzymatic degradation. This flexible protective layer maintains the stability of the delivered components without significantly increasing system complexity
Data Source
AI summary
The present invention relates to a CRISPR nanocomplex for nonviral genome editing, a method for preparing the same, and the like. The CRISPR nanocomplex for nonviral genome editing of the present invention has a size of several nanometers to several microns, enables intracellular delivery without external physical stimulation, and can be utilized for genome editing through nonviral routes with respect to target genes of cells. As a result, when used for preparation of animal model, microbiological engineering, cell engineering for disease treatment, or formulations for biological administration, the CRISPR Nanocomplex shows high intracellular delivery and gene editing efficiency, and can minimize problems, such as nonspecific editing, gene mutation, and induction of cytotoxicity and biotoxicity.