Trigger Oligo HCR Probe for Specific Signal Amplification
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Solution Overview
Problem
Existing hybridization chain reaction (HCR) techniques face issues with non-specific binding and background amplification due to exposed HCR initiators, leading to reduced sample penetration and increased stickiness of antibody probes, which results in poor target detection and signal amplification.
Innovation Solution
Incorporating a trigger oligo that binds to a structured nucleic acid probe to expose the HCR initiator only when it selectively hybridizes to the target, reducing non-specific binding and enhancing signal amplification by using modified HCR hairpins with sequestered initiators that are activated by the oligo, thereby minimizing background noise.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If HCR initiator is exposed on antibody probes to enable signal amplification, then signal amplification is improved, but non-specific binding and background amplification increase
Solution Approach 1:
The HCR initiator is extracted from the antibody probe structure and separated into a distinct trigger oligo component. This allows the antibody probe to function without the harmful exposed initiator, while the initiator can only be activated when specifically bound to the target, thereby eliminating non-specific binding and background amplification while preserving signal amplification capability.
Solution Approach 2:
The trigger oligo containing the HCR initiator is designed to bind to the target molecule first, and only then activate the HCR signal amplification. This preliminary binding action ensures that signal amplification occurs only at the specific target location, preventing background amplification and non-specific binding while maintaining high signal amplification efficiency.
2Measurement precision
If antibody probes are made sticky to enhance target binding, then target detection sensitivity is improved, but sample penetration ability deteriorates
Solution Approach 1:
The harmful 'stickiness' property is extracted from the antibody probe by removing the exposed HCR initiator that caused non-specific binding. This allows the probe to penetrate samples more effectively without sacrificing target detection sensitivity, as the specific binding capability is maintained through the trigger oligo mechanism.
Solution Approach 2:
The trigger oligo acts as an intermediary that mediates between the antibody probe and the HCR initiator. This intermediary ensures that the probe does not need to be sticky to achieve high target detection sensitivity, as the trigger oligo provides the necessary binding specificity while allowing the probe to maintain better sample penetration ability.
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
This approach improves the specificity and sensitivity of target detection by reducing non-specific binding and background amplification, allowing for deeper sample penetration and increased signal generation per target molecule, while maintaining high signal-to-background ratios.
Implementation Method 1
a trigger oligo, wherein the trigger oligo binds to the structured nucleic acid region
Implementation Method 2
which changes conformation to expose the initiator
Implementation Method 3
Hybridization chain reaction is a method for the triggered self-assembly of nucleic acid molecules starting from metastable hairpin monomers
Implementation Method 4
triggered self-assembly of nucleic acid molecules
Implementation Method 5
amplification via polymerization of fluorophore-labeled hairpins
Data Source
Figure 1A
Figure 1B
Figure 1C
AI summary
The present disclosure relates to methods involving HCR reactions that involve using trigger oligos to activate probes that initiate HCR