DNA Aptamer for Methylated DNA Detection
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Solution Overview
Problem
Current methods for detecting methylated DNA fragments, such as Methylated DNA Immunoprecipitation (MeDIP), face challenges like high cost and non-specific binding due to the use of antibodies, lacking effective alternatives for capturing and detecting these fragments.
Innovation Solution
Development of a DNA aptamer specifically binding to methylated DNA fragments, selected from sequences like SEQ ID NO:7 and SEQ ID NO:9, which forms a complex with the aptamer and can be used for detection and amplification, along with a method involving SELEX technology to select aptamers that bind to methylated DNA fragments under specific conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If MeDIP uses monoclonal antibodies to capture methylated DNA fragments, then detection capability is achieved, but cost increases and non-specific binding occurs
Solution Approach 1:
The patent replaces antibodies with DNA aptamers that copy the binding function. The aptamers are selected through SELEX to specifically bind methylated DNA fragments, replicating the capture capability of antibodies while avoiding protein-related non-specific binding issues.
Solution Approach 2:
The patent changes the molecular type from protein (antibody) to nucleic acid (aptamer). This parameter change fundamentally alters the binding characteristics, reducing non-specific binding while maintaining detection capability through sequence-specific recognition of methylated DNA.
2Measurement precision
If MeDIP uses monoclonal antibodies to capture methylated DNA fragments, then detection capability is achieved, but cost increases
Solution Approach 1:
The patent employs DNA aptamers which are chemically synthesized and generally less expensive than monoclonal antibodies. Aptamers can be produced through in vitro selection and chemical synthesis, avoiding the costly animal immunization and hybridoma production processes required for antibodies.
Solution Approach 2:
The aptamer system copies the functional role of antibodies but uses a cheaper molecular platform. The SELEX process generates aptamers that perform the same capture function at lower material and production costs.
3Measurement precision
If DNA aptamer is used to bind methylated DNA fragment, then specificity and sensitivity are improved, but selection process complexity increases
Solution Approach 1:
The SELEX process is a self-organizing in vitro evolution system where the aptamer library automatically selects high-affinity binders through iterative binding and amplification cycles. The system self-optimizes without requiring complex external intervention, achieving high specificity through natural selection principles.
Solution Approach 2:
The patent performs preliminary selection of aptamers with high affinity and specificity for methylated DNA through multiple rounds of SELEX before application. This preliminary action ensures that the selected aptamers possess the required binding characteristics, simplifying subsequent experimental procedures.
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 DNA aptamer demonstrates high specificity and sensitivity in capturing and detecting methylated DNA fragments, reducing costs and non-specific binding issues, enabling effective analysis of epigenetic regulation and potential therapeutic targets.
Implementation Method 1
a DNA aptamer specifically binding to a methylated DNA fragment
Data Source
AI summary
A DNA aptamer specifically binding to a methylated DNA fragment includes a nucleotide sequence selected from the group consisting of SEQ ID NO:7 and SEQ ID NO:9. In addition, a method for detecting a methylated DNA fragment in a sample and a method for selecting a DNA aptamer specifically binding to the methylated DNA fragment are also disclosed.