Helz2 Knockout Mouse Model via CRISPR/Cas9 for Tumor Therapy
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Solution Overview
Problem
There is a lack of research and application of Helicase with zinc finger 2 (Helz2) knockout mouse models in the anti-tumor aspect, and no corresponding research or application using the CRISPR/Cas9 system for targeting Helz2 or its expressed product in tumor therapy.
Innovation Solution
A method is developed to establish a Helz2 knockout mouse model using the CRISPR/Cas9 technique, involving the use of specific sgRNA sequences for fertilized mouse eggs, resulting in a systemic Helz2 knockout homozygous mouse model, and a vector containing target sequences to interfere with Helz2 expression, which can be used to treat or prevent tumors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If CRISPR/Cas9 technique is used to establish Helz2 knockout mouse model, then the precision of gene knockout is improved, but the complexity of the experimental procedure increases
Solution Approach 1:
The patent uses sgRNA as an intermediary molecule that guides the Cas9 enzyme to the specific Helz2 gene target sequence. This mediator enables precise gene knockout by facilitating the binding of Cas9 to the correct genomic location, resolving the contradiction between precision and complexity by providing a specific molecular mechanism for targeted editing.
2Object-affected harmful factors
If Helz2 knockout is performed to inhibit tumor growth, then the anti-tumor effect is improved, but the risk of off-target effects increases
Solution Approach 1:
The patent designs sgRNA sequences that are highly specific to the Helz2 gene, ensuring that the gene knockout effect is localized precisely to the intended target. This local specificity minimizes off-target effects while maintaining the desired anti-tumor effect, resolving the contradiction between therapeutic efficacy and safety.
3Object-affected harmful factors
If vector with target sequence is used to interfere with Helz2 expression, then the tumor treatment efficacy is improved, but the complexity of drug delivery system increases
Solution Approach 1:
The patent extracts the essential functional element (target sequence) from the complex vector system and identifies it as the key active component. By focusing on the target sequence itself (SEQ ID NOs: 1-7) as the therapeutic agent, the patent simplifies the drug delivery concept while maintaining treatment efficacy, resolving the contradiction between therapeutic effect and system complexity.
Data Source
AI summary
The present disclosure provides a vector containing a target sequence that targets and interferes with Helicase with zinc finger 2 (Helz2), the target sequence comprising one or more of the following sequences:SEQ ID No. 5:5′-GCTATCAAGTCTGTCACTACT-3′;SEQ ID No. 6:5′-GCACGATGCTGTATGGCTTTG-3′;SEQ ID No. 7:5′-GGGCCTCATTGACACTCAAAG-3′.