Self-Replicating Reporter Nucleic Acid for Methylation Detection
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Solution Overview
Problem
Current methods for obtaining epigenetic information from cells are complex and inefficient, particularly in detecting methylation status, which is crucial for health condition assessment and disease diagnosis.
Innovation Solution
A self-replicating reporter nucleic acid construct is introduced into cells, which transcribes and reports the methylation status of specific genome sequences by producing a detectable signal, allowing for easy detection of epigenetic information through signal presence or quantity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods (bisulfate treatment PCR, sequencing) are used to detect DNA methylation, then methylation status can be determined, but the process is complex and inefficient
Solution Approach 1:
The patent introduces a self-replicating vector as an intermediary system that mediates between the cell's endogenous DNA methylation machinery and the detection system. The vector contains a reporter gene under the control of a promoter whose activity reflects the methylation status of the target genomic region, thereby translating complex epigenetic information into a simple detectable signal without requiring complex bisulfate treatment or sequencing procedures
Solution Approach 2:
The patent replaces complex mechanical and chemical processing steps (bisulfate treatment, PCR amplification, sequencing) with a biological reporting system. The self-replicating vector utilizes the cell's own transcriptional and replication machinery to generate a detectable signal that directly reports methylation status, substituting cumbersome laboratory procedures with an in-cell biological assay
2Loss of information
If conventional methylation detection methods are used, then epigenetic information can be obtained, but the process is time-consuming and inefficient
Solution Approach 1:
The self-replicating vector is designed with pre-assembled regulatory elements (promoter, reporter gene, origin of replication) that are ready to immediately interact with the cell's methylation machinery upon introduction. The vector's promoter region is configured to respond directly to the methylation status of the target genome, eliminating the need for time-consuming sample preparation and allowing immediate detection of epigenetic information
Solution Approach 2:
The self-replicating vector utilizes the cell's own transcriptional and replication machinery to generate the detection signal. The vector autonomously replicates and expresses the reporter gene based on the local methylation environment, eliminating the need for external processing steps and enabling efficient, self-contained epigenetic information acquisition within the living cell
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 method simplifies the acquisition of epigenetic information, enabling precise detection of methylation status and potential disease markers, such as cancer, by utilizing a self-replicating vector that produces a detectable signal indicative of methylation levels.
Implementation Method 1
The methylation reaction of genomic DNA is that a methyl group is added to the cytosine on a genome by DNA methyltransferase in a cell. When a genome is replicated at the time of cell division, a methyl group is added to the cytosine in the reproduced genome at the same position as that of the genome before the replication.
Implementation Method 2
detecting the presence/absence of a signal produced in the subject cell and/or a size thereof
Data Source
Figure 1~2
Figure 3
Figure 4(a)~4(c)
AI summary
According to an embodiment, a method of obtaining epigenetic information of a cell is provided. The method includes introducing a reporter nucleic acid construct in a nuclear of a subject cell, self-replication of the reporter nucleic acid construct, detecting the presence/absence of a signal produced in the subject cell and/or a size thereof, and obtaining epigenetic information of the subject cell based on the result obtained. The reporter nucleic acid construct is characterized by transcribing the state of modification in a specific sequence on the genome of the subject cell onto a corresponding sequence thereof by substitution of a functional group during the self-replication in the nuclear of the subject cell, and producing a detectable signal depending on the state of presence of the transcribed functional group.