Cross-linked Nucleic Acid Complex for Nuclease Resistance
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Solution Overview
Problem
Existing methods for nucleic acid hybridization in living organisms face challenges due to instability caused by nucleases, high synthesis costs of modified nucleic acids, and fluctuating stability of complementary double-stranded structures with external factors like temperature, ionic strength, or pH.
Innovation Solution
A nucleic acid complex comprising a single-stranded nucleic acid and a cross-linked double-stranded nucleic acid, where the cross-linked double-stranded nucleic acid is linked to the 5′ or 3′ end of the single-stranded nucleic acid, providing enhanced stability through covalent bonds and a cross-linking reagent, allowing for stable hybridization with target nucleic acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If chemically modified nucleic acids (LNAs, 2′-O-methyl nucleotides) are used to improve stability, then nucleic acid stability in hybridization is improved, but synthesis cost increases significantly
Solution Approach 1:
The patent uses unmodified nucleic acid sequences that are cheaper to synthesize, combined with a disposable cross-linking approach where the cross-linking reagent forms stable bonds between complementary strands, eliminating the need for expensive modified nucleic acids while maintaining stability
Solution Approach 2:
The patent creates a composite structure by forming cross-linked double-stranded nucleic acid complexes using standard nucleic acids combined with cross-linking reagents (such as formaldehyde, glutaraldehyde, or carbodiimide chemistry), achieving stability comparable to modified nucleic acids without the high synthesis cost
2Stability of the object's composition
If complementary double-stranded nucleic acid structures are formed to improve nuclease resistance, then stability is improved, but the structure becomes sensitive to external factors (temperature, ionic strength, pH) causing dissociation
Solution Approach 1:
The patent applies cross-linking reagents to the complementary double-stranded nucleic acid structures before they are exposed to varying environmental conditions or nucleases. The cross-links are formed in advance to prevent dissociation, counteracting the destabilizing effects of temperature, ionic strength, or pH changes before they can cause structural failure
Solution Approach 2:
The cross-linking reaction is performed as a preliminary step to stabilize the double-stranded structure before introducing it to the biological environment. This preliminary cross-linking action ensures that the structure maintains its integrity under subsequent environmental variations and nuclease exposure
3Stability of the object's composition
If multiple nucleic acids are modified in the oligonucleotide sequence, then stability is improved, but synthesis cost increases
Solution Approach 1:
The patent replaces expensive modified nucleic acids with unmodified sequences that can be synthesized more efficiently using standard protocols, accepting that the unmodified sequences would normally be less stable, but compensating for this through the cross-linking mechanism
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 nucleic acid complex achieves stable hybridization with target nucleic acids, reduces nuclease resistance, and lowers synthesis costs, maintaining stability across varying environmental conditions.
Implementation Method 1
a cross-linked double-stranded nucleic acid comprising a first nucleic acid strand linked to at least one of the 5′ end and the 3′ end of the single-stranded nucleic acid and a second nucleic acid strand comprising a base sequence that is completely or sufficiently complementary to the first nucleic acid strand
Data Source
AI summary
A nucleic acid complex includes a single-stranded nucleic acid and a cross-linked double-stranded nucleic acid including the first nucleic acid strand linked to at least one of the 5′ end and the 3′ end of the single-stranded nucleic acid and the second nucleic acid strand including a base sequence that is completely or sufficiently complementary to the first nucleic acid strand.