shTACE Gene/Carrier Complex for Inflammatory Disease Treatment
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Solution Overview
Problem
Current methods for obtaining shRNA targeting TACE (shTACE) have low yield and purity issues due to the use of Miniprep, which requires repeated processes and does not effectively remove endotoxins, and shTACE has low stability, making it challenging for efficient gene delivery in gene therapy.
Innovation Solution
Development of a gene/carrier complex using modified shTACE sequences and nonviral carriers like PAs-s and 8D16R, which form a stable complex for efficient intracellular delivery, overcoming yield and stability issues by using a weight ratio optimization and electrostatic attraction for enhanced therapeutic effect.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If Miniprep is used to obtain shTACE, then the difficulty of amplification is reduced, but the gene yield is much lower and endotoxins are not removed
Solution Approach 1:
The patent modifies the shTACE sequence parameters (specifically the hairpin loop structure and flanking sequences) to optimize for Maxiprep conditions, achieving high yield while maintaining stability. This parameter optimization allows the use of Maxiprep instead of Miniprep, resolving the contradiction between ease of manufacture and gene yield.
Solution Approach 2:
The patent creates a modified version of the shTACE sequence that is optimized for high-yield amplification. By copying and modifying the original sequence with improved features (such as enhanced hairpin structure and stable flanking regions), the patent achieves both high yield and stability without requiring repeated Miniprep procedures.
2Ease of manufacture
If Miniprep is used to obtain shTACE, then the difficulty of amplification is reduced, but additional experiments are required to remove endotoxins
Solution Approach 1:
By optimizing the shTACE sequence parameters for Maxiprep conditions, the patent enables a single-step high-yield amplification process that inherently removes endotoxins. This eliminates the need for additional endotoxin removal experiments, reducing overall process complexity while maintaining ease of manufacture.
3Quantity of substance
If Maxiprep is used to obtain shTACE, then the gene yield is improved and endotoxins are removed, but the amplification degree is very low for difficult sequences
Solution Approach 1:
The patent optimizes the shTACE sequence parameters (hairpin structure, flanking sequences, and overall composition) to be compatible with Maxiprep conditions. This parameter optimization enables high-yield amplification with good stability, resolving the contradiction between gene yield and amplification difficulty.
Solution Approach 2:
The patent creates a composite shTACE construct combining optimized hairpin structure with stable flanking sequences. This composite design enhances both amplification efficiency and stability, allowing Maxiprep to achieve high yield without the amplification difficulties that plague simpler sequences.
4Reliability
If shTACE is used as a gene therapy agent, then the inflammatory disease treatment effect is achieved, but the stability in the human body is low
Solution Approach 1:
The patent modifies the shTACE sequence parameters to enhance stability in the human body while maintaining therapeutic effect. Specific changes include optimizing the hairpin loop structure, adding stable flanking sequences, and adjusting the overall sequence composition to resist degradation, thereby resolving the contradiction between therapeutic effect and stability.
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 gene/carrier complex achieves effective inhibition of TACE expression, reducing inflammatory cytokines like TNF-α, and demonstrates improved yield, stability, and delivery efficiency, making it suitable for treating inflammatory diseases.
Implementation Method 1
nonviral carriers like PAs-s and 8D16R, which form a stable complex for efficient intracellular delivery, overcoming yield and stability issues by using a weight ratio optimization and electrostatic attraction for enhanced therapeutic effect
Data Source
AI summary
Disclosed is a gene/carrier complex for preventing or treating inflammatory diseases, including tumor necrosis factor-α converting enzyme (TNF-α converting enzyme, TACE) shRNA and a nonviral gene carrier, wherein the nonviral gene carrier includes an acetate of disulfide-linked poly(oligo-arginine) or a TFA salt of poly(oligo-aspartic acid)poly(oligo-arginine).