Duplex Nucleic Acid Label for Bright Fluorescence Without Self-Quenching
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Solution Overview
Problem
The incorporation of numerous individual dyes within markers leads to self-quenching, significantly diminishing fluorescence intensity and complicating quantitative analysis, especially in high-resolution studies.
Innovation Solution
A label comprising a first and second nucleic acid strand forming a duplex structure with optically detectable labelling moieties attached, and a binding molecule to enhance rigidity, preventing aggregation and maintaining native optical properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If numerous individual dyes are incorporated within markers, then sensitivity and detection capability are improved, but self-quenching occurs which diminishes fluorescence intensity and complicates quantitative analysis
Solution Approach 1:
The label is segmented into two separate nucleic acid strands that form a duplex structure, with labelling moieties attached only to the first strand. This spatial segmentation prevents aggregation while maintaining high dye density for sensitive detection.
Solution Approach 2:
The second nucleic acid strand acts as an intermediary that binds to the first strand to form a rigid duplex structure. This intermediary backbone maintains fixed spacing between labelling moieties, preventing self-quenching while preserving fluorescence intensity for quantitative analysis.
2Illumination intensity
If numerous individual dyes are incorporated within markers, then bright emission is achieved, but aggregation of labelling moieties occurs which changes their native optical properties
Solution Approach 1:
The duplex structure creates locally rigid regions that maintain fixed spatial relationships between labelling moieties. This local structural quality prevents aggregation and preserves the native optical properties of each dye while allowing high concentration for bright emission.
Solution Approach 2:
The patent changes the structural parameter of the nucleic acid backbone from single-stranded to duplex structure, increasing rigidity and persistence length. This parameter change maintains labelling moiety spacing and prevents aggregation, preserving optical properties while enabling high dye density for bright fluorescence.
3Ease of manufacture
If flexible nucleic acid strands are used, then ease of manufacture is improved, but rigidity is insufficient which allows aggregation of labelling moieties
Solution Approach 1:
Two flexible single-stranded nucleic acids merge to form a rigid duplex structure through complementary base pairing. This merging maintains ease of manufacture since both strands are independently synthesizable, while the combined structure provides the necessary rigidity to prevent labelling moiety aggregation.
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 label achieves bright and efficient emission of labelling moieties, enabling unambiguous identification and analysis of target analytes with high sensitivity and specificity.
Implementation Method 1
The first nucleic acid strand and the second nucleic acid strand are configured to form at least one duplex structure
Implementation Method 2
at least one binding molecule configured to bind to the at least one duplex structure
Implementation Method 3
The label enables bright emission of the plurality of labelling moieties, in particular with native optical properties of the labelling moieties, at a high efficiency
Data Source
AI summary
A label (100) for analysing a biological sample is provided. The label comprises a first nucleic acid strand (102) and a second nucleic acid strand (106). The first nucleic acid strand (102) and the second nucleic acid strand (106) are configured to form at least one duplex structure. The label further comprises a first plurality of optically detectable labelling moieties (104) attached to the first nucleic acid strand (102). The label further comprises at least one binding molecule (108) configured to bind to the at least one duplex structure. In further aspects a marker (200) and a method for detecting target analytes (112, 300, 302) are provided.