Primate Liver Cancer Model via Hepatic Portal CRISPR Injection
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Solution Overview
Problem
The construction of primate disease models using CRISPR technology is hindered by the immaturity of embryonic stem cell and somatic cell nuclear transplantation methods in non-human primates, resulting in long cycles and high costs, limiting the effectiveness of disease modeling.
Innovation Solution
A method involving the construction of a sgRNA expression plasmid using a gRNA oligonucleotide and pX330 plasmid, followed by direct injection into the hepatic portal vein of a primate animal to induce gene mutations in liver cells, specifically targeting the p53 gene for rapid tumor model development.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If embryonic stem cell or somatic cell nuclear transplantation technology is used to construct primate disease models, then precise gene modification can be achieved, but the construction cycle becomes long and cost increases
Solution Approach 1:
The patent changes the delivery route parameter from traditional embryonic stem cell transplantation to direct hepatic portal vein injection of CRISPR-Cas9 components. This parameter change enables somatic cell editing in adult primates, dramatically shortening the construction cycle from years to months while maintaining gene modification precision through targeted delivery of sgRNA and Cas9 plasmids
Solution Approach 2:
The patent uses the hepatic portal vein as an intermediary delivery route to transport CRISPR-Cas9 components directly to liver tissue. This intermediary pathway bypasses the need for embryonic stem cell manipulation and nuclear transplantation, enabling direct in vivo gene editing and significantly reducing the time and complexity of model construction
2Manufacturing precision
If traditional gene targeting technology based on embryonic stem cells is used, then specific genes can be precisely modified, but the method complexity and cost increase
Solution Approach 1:
The patent extracts the essential gene editing function (CRISPR-Cas9 system) from the complex embryonic stem cell transplantation process. By delivering only the necessary components (sgRNA expression plasmid and Cas9 plasmid) directly into the hepatic portal vein, the method eliminates the need for embryonic stem cell culture, manipulation, and transplantation infrastructure, greatly simplifying the overall process while maintaining editing precision
Solution Approach 2:
The patent uses plasmid-based CRISPR-Cas9 delivery as a simplified copy or alternative version of the gene editing function. Instead of requiring the full embryonic stem cell system, the plasmids serve as self-contained copies of the gene editing machinery that can be delivered directly, reducing device and procedural complexity while achieving the same gene modification outcome
3Productivity
If CRISPR technology is applied at the somatic cell level in primates, then the model construction cycle can be shortened, but the technology availability and effectiveness are limited
Solution Approach 1:
The hepatic portal vein serves as an intermediary delivery route that overcomes the limitation of CRISPR technology availability in primates. This route enables direct delivery of CRISPR components to liver tissue, bypassing the need for mature primate embryonic stem cell transplantation technology and making somatic cell editing feasible in this species
Solution Approach 2:
The patent changes the delivery parameter from traditional in vitro embryonic stem cell manipulation to direct in vivo injection via hepatic portal vein. This parameter change transforms CRISPR technology from being unavailable or difficult to apply in primates to being readily implementable, thereby improving reliability and enabling productive model construction
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 enables the rapid and efficient construction of primate disease models by directly injecting the sgRNA expression plasmid into primate liver tissue, achieving a high mutation rate and inducing liver cancer, thus simulating human disease conditions effectively.
Implementation Method 1
When exogenous DNA invades, CRISPR-RNA directs the CAS protein to perform a specific splicing and generates double strand breaks (DSBs) at the DNA target sites
Implementation Method 2
a biolistic method for delivering a plasmid into a liver cell
Data Source
AI summary
Provided is a primate disease model construction method based on fast gene edition, which including (a) constructing a sgRNA expression plasmid by using a gRNA oligonucleotide and a pX330 plasmid; (b) injecting the sgRNA expression plasmid prepared in step (a) into a hepatic portal vein of a primate animal by using a biopsy needle until liver cells become cancerous for obtaining a primate disease model. The sgRNA expression plasmid constructed by the gRNA oligonucleotide and pX330 plasmid can be directly injected into the primate liver tissue, so as to construct a tumor model rapidly.