Covalent Chromosome Matrix Attachment for 3D Structure Preservation

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Solution Overview

Problem

Current methods for visualizing and analyzing cellular structures, such as fluorescence in situ hybridization (FISH), face challenges in preserving the three-dimensional structure and spatial orientation of nucleic acids within cells, limiting the accuracy and detail of chromosome visualization.

Innovation Solution

The method involves covalently attaching chromosomes to a matrix within a cell by modifying nucleotides with a matrix attachment moiety and using oligonucleotides, such as oligopaints, to represent the chromosome structure, allowing for the preservation of three-dimensional nucleic acid sequences and spatial orientation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If FISH is applied to fixed cell or tissue samples, then chromosome positioning and spatial orientation can be revealed, but the three-dimensional structure and spatial orientation of nucleic acids are not preserved with sufficient accuracy

Engineering Contradiction:
Improvechromosome visualization accuracyVSAvoidthree-dimensional structure preservation
Core Design Contradiction:
Measurement precisionVSStability of the object's composition

Solution Approach 1:

The patent applies preliminary action by first modifying nucleotides within chromosomes to include matrix attachment moieties before fixing the cell samples. This pre-modification ensures that when the matrix is subsequently introduced and attached, the three-dimensional structure is already prepared for accurate preservation and visualization, resolving the contradiction between measurement precision and structural stability.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent introduces a matrix as an intermediary substance that mediates between the chromosome and the visualization process. The matrix attachment moiety on modified nucleotides binds to this matrix, creating a stable framework that preserves three-dimensional structure while enabling accurate chromosome positioning and spatial orientation to be revealed, thus resolving the technical contradiction.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Reliability

If nucleotides are modified to include matrix attachment moiety, then chromosome can be covalently attached to matrix, but the complexity of the procedure increases

Engineering Contradiction:
Improvechromosome attachment stabilityVSAvoidprocedure complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent applies segmentation by dividing the attachment process into distinct functional components: a matrix attachment moiety that is incorporated into the nucleotide structure, a separate matrix substance that is introduced into the cell, and the covalent bond that forms between them. This segmentation allows each component to be optimized independently, improving attachment reliability while making the overall procedure more manageable despite the added complexity.

Inventive Principle:
Principle #1Segmentation

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 enables precise visualization and modeling of chromosome structures within cells, maintaining their native three-dimensional shape and relative positions, enhancing the accuracy of genetic analysis and gene expression studies.

Implementation Method 1

hybridizing to the chromosome a plurality of oligonucleotides

Methodology Applied
Scientific EffectHybridization:

Implementation Method 2

covalently attaching a chromosome within a cell to a matrix including modifying a plurality of nucleotides within the chromosome to include a matrix attachment moiety

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Data Source

PatentEP3445875B1Methods for attaching cellular constituents to a matrix
Publication Date: 2023.01.18 PRESIDENT & FELLOWS OF HARVARD COLLEGE
  • EP3445875B1 patent drawingFigure 1
  • EP3445875B1 patent drawingFigure 2
  • EP3445875B1 patent drawingFigure 3A~3E

AI summary

The present invention relates to a method of covalently attaching a chromosome within a cell to a matrix including modifying a plurality of nucleotides within the chromosome to include a matrix attachment moiety wherein the chromosome contacts the matrix, and attaching the matrix attachment moiety of the plurality of nucleotides to the matrix, thereby attaching the chromosome to the matrix.