Nucleic Acid Cleavage Kit for Gene Therapy
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Solution Overview
Problem
Conventional nucleic acid cleavage tools, such as restriction enzymes, lack the necessary sequence specificity to effectively recognize and cleave longer nucleic acid sequences, which is a limitation in applications like gene therapy and genetic analysis, especially for sequences over 8 base pairs.
Innovation Solution
A nucleic acid cleavage kit comprising a carrier, an oligonucleotide, and a nucleic acid cleavage agent, where the oligonucleotide is bound to the carrier to recognize and the agent is bound to cleave the target nucleic acid, enabling site-specific cleavage with high sequence specificity by forming double or triple helix structures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional restriction enzymes are used for nucleic acid cleavage, then the cleavage process is simple and well-established, but the sequence recognition capability is limited to 8 base pairs or less
Solution Approach 1:
The patent combines three functional components into a unified cleavage system: (1) oligonucleotide for sequence recognition, (2) carrier for stabilization and delivery, and (3) nucleic acid cleavage agent for cutting. This merging allows the system to achieve recognition of 10-15 base pairs or more while maintaining functional effectiveness, resolving the contradiction between enhanced sequence specificity and system complexity.
Solution Approach 2:
The invention creates a composite cleavage system where chemically distinct components (oligonucleotide, carrier molecule, and cleavage agent) are assembled together through specific binding interactions. The oligonucleotide portion provides sequence-specific recognition capability beyond 8 base pairs, while the carrier and cleavage agent provide functional stability and cutting activity, respectively. This composite approach enables extended sequence recognition without sacrificing cleavage efficiency.
2Reliability
If oligonucleotides are used alone as probes, then high sequence-specific recognition is achieved, but they cannot perform cleavage function
Solution Approach 1:
The patent merges the recognition function of oligonucleotides with the cleavage function of nucleic acid cleavage agents by assembling them into a single multifunctional system. The oligonucleotide portion binds to the target sequence with high specificity (10-15 base pairs or more), while the attached cleavage agent performs the actual cutting. This combination allows the system to simultaneously achieve high recognition reliability and cleavage versatility.
Solution Approach 2:
The oligonucleotide-cleavage agent conjugate serves multiple functions: (1) sequence-specific recognition through oligonucleotide binding, (2) stabilization through carrier interaction, and (3) cleavage through the attached nucleic acid cleavage agent. This multi-functionality resolves the contradiction between specialized recognition capability and overall functional versatility.
3Measurement precision
If recognition length is extended beyond 8 base pairs, then sequence specificity is improved, but conventional restriction enzymes become incompetent
Solution Approach 1:
The invention changes the fundamental parameters of the cleavage system by replacing conventional restriction enzymes with a modular oligonucleotide-cleavage agent conjugate. This parameter change allows the system to achieve recognition of 10-15 base pairs or more while maintaining effective cleavage, as the cleavage agent's activity is activated by its binding to the extended oligonucleotide sequence rather than relying on traditional enzyme recognition sites.
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 kit provides precise cleavage of nucleic acids with recognition capabilities beyond 10 nucleotide units, enhancing the scope of nucleic acid cleavage applications, reducing side effects, and expanding the utility in gene therapy and genetic analysis.
Implementation Method 1
The first end of the oligonucleotide is bound with the carrier to recognize at least partial sequence of the target nucleic acid
Data Source
AI summary
A nucleic acid cleavage kit is used to cleave a target nucleic acid. The nucleic acid cleavage kit includes a carrier, an oligonucleotide, and a nucleic acid cleavage agent. The oligonucleotide recognizes at least partial sequence of the target nucleic acid. Then, the nucleic acid cleavage agent cleaves the target nucleic acid. A nucleic acid cleavage detection apparatus including the nucleic acid cleaving kit and a gene therapy by administering the nucleic acid cleavage kit are also disclosed.