Caged Nucleic Acid Fragment for Single-Cell Analysis
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Solution Overview
Problem
Current methods for analyzing the base sequence of target nucleic acids at the level of one cell or sub-cell require labor-intensive processes like laser microdissection, making them inefficient and complex.
Innovation Solution
A nucleic acid fragment with polymerase and transposase binding sequences containing caged nucleotide residues is used, which is irradiated with active energy rays to eliminate caging groups, allowing for localized binding and insertion near DNA-binding proteins, enabling gene amplification and analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If laser microdissection is used to analyze target nucleic acids at the level of one cell, then measurement precision is improved, but device complexity and ease of operation deteriorate
Solution Approach 1:
The patent replaces the mechanical laser microdissection system with a chemical approach using caged nucleotide residues that can be activated by light to enable polymerase binding and DNA amplification directly in situ, eliminating the need for complex mechanical cutting and manipulation equipment
Solution Approach 2:
The patent introduces caged nucleotide residues as an intermediary substance that mediates between the target DNA and polymerase binding. These residues remain inactive until activated by light irradiation, at which point they enable specific polymerase binding and subsequent DNA amplification, providing a simple chemical mechanism for single-cell analysis
2Measurement precision
If laser microdissection is used to isolate target cells, then measurement precision is improved, but ease of operation deteriorates
Solution Approach 1:
The patent replaces manual laser microdissection operations with a simple chemical activation process. Researchers只需 irradiate the sample with light at the desired location to activate caged nucleotide residues, which then automatically enable polymerase binding and DNA amplification, eliminating complex manual manipulation
Solution Approach 2:
The caged nucleotide residues perform self-service by automatically activating upon light irradiation to enable polymerase binding and DNA amplification without requiring external manipulation or additional reagents, simplifying the operational process while maintaining single-cell analysis precision
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 simplifies the analysis of target nucleic acids by allowing for localized, efficient insertion and amplification at the level of one cell or sub-cell, overcoming the complexity of existing methods.
Implementation Method 1
a step of irradiating the nucleic acid fragment with active energy rays and eliminating a caging group from the caged nucleotide residue
Data Source
AI summary
A nucleic acid fragment including a polymerase binding sequence and a transposase binding sequence, in which the polymerase binding sequence or the transposase binding sequence contains a caged nucleotide residue; a specific binding substance; a method for binding a polymerase or a transposase to the nucleic acid fragment; a method for inserting the nucleic acid fragment in a vicinity of a binding region of a DNA-binding protein bound to a DNA molecule; and a method for gene-amplifying the DNA molecule into which the nucleic acid fragment is inserted are provided.