Tagged Split G-Quadruplexes for Stable Nucleic Acid Capture
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Solution Overview
Problem
Split G-quadruplexes used for nucleic acid detection exhibit low target sensitivity due to weaker peroxidase activity and higher inactivation rates, limiting their use as effective nucleic acid detection agents compared to horseradish peroxidase (HRP).
Innovation Solution
Associating tags with split G-quadruplexes to enhance their binding affinity and specificity for target nucleic acids, allowing for stable interaction and detection or capture of nucleic acids, mimicking the binding properties of antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If split G-quadruplexes are used for nucleic acid detection, then they can bind to target nucleic acids, but their peroxidase activity is weaker and inactivation rates are higher, resulting in low target sensitivity
Solution Approach 1:
The patent introduces an intermediary detection system where split G-quadruplexes bind to target nucleic acids but do not directly provide the detection signal. Instead, they recruit HRP-containing nanoparticles that serve as the actual detection agents. This mediator approach allows the G-quadruplex to maintain its binding function while overcoming its weak peroxidase activity through the use of a more effective enzymatic mediator.
2Adaptability or versatility
If split G-quadruplexes are used as detection agents, then they can specifically bind to target sequences, but their catalytic activity is insufficient compared to horseradish peroxidase
Solution Approach 1:
The patent merges the advantages of two different systems: the high binding specificity of split G-quadruplexes and the strong catalytic activity of HRP-containing nanoparticles. By combining these two components into a unified detection system, the invention achieves both high specificity (from the G-quadruplex binding) and high power (from the HRP catalysis), resolving the contradiction between adaptability and power.
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 tagged split G-quadruplexes demonstrate high binding affinity and specificity, enabling stable interaction with target nucleic acids without requiring peroxidase activity, thus improving detection and capture methods, potentially surpassing the limitations of existing split G-quadruplex applications.
Implementation Method 1
G-quadruplexes are structures formed in nucleic acids by sequences that are rich in guanine. Four guanine bases can associate through Hoogsteen hydrogen bonding to form a square planar structure called a guanine tetrad
Implementation Method 2
The quadruplex structure is further stabilized by the presence of a cation, which sits in a central channel between each pair of tetrads
Implementation Method 3
With addition of oxidizing agent H2O2, the G-quarduplex-hemin complex is capable of oxidizing a variety of substrates, including colorimetric and chromogenic substrates (ex. DAB, ABTS)—and chemiluminescent substrates (ex. luminol)—used in peroxidase assays
Implementation Method 4
the G-quarduplex-hemin complex is capable of oxidizing a variety of substrates
Data Source
AI summary
Methods of using split G-quadruplexes associated with functional tags for associating said tags to target nucleic acids. Methods include use of split G-quadruplexes associated with detection tags for the detection of target nucleic acids, and use of split G-quadruplexes associated with capture tags for detection or capture of target nucleic acids.