CRISPR/Cas Gene Editing for Aneuploidy Correction

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Solution Overview

Problem

Current methods for correcting aneuploidies and frame shift mutations in embryos are limited by mosaicism and the physical separation of maternal and paternal genomes during early development, making precise and efficient correction challenging.

Innovation Solution

The use of RNA-guided endonucleases, specifically CRISPR/Cas systems, to introduce targeted double-stranded breaks in embryos, allowing for the precise elimination of extra chromosomes or correction of frame shift mutations by directing the endonuclease to specific sites within the genome, thereby correcting aneuploidies and frame shift mutations in a non-mosaic manner.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If embryo selection and elimination by aneuploidy testing is performed in IVF clinics, then aneuploidies are detected and eliminated, but fertility rates are reduced due to loss of affected embryos

Engineering Contradiction:

Improveaneuploidy detection accuracyVSAvoidfertility rates

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The invention converts the harmful effect of aneuploidy (chromosomal abnormality) into a beneficial outcome by using the presence of the extra chromosome itself as the target for correction. RNA-guided endonucleases are designed to specifically recognize and eliminate the extra chromosome in trisomic embryos, transforming the problematic aneuploid condition into a correctable defect that can be precisely removed without affecting normal chromosomes.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

2Manufacturing precision

If CRISPR-based gene editing is used to correct mutations, then precise targeted correction is achieved, but mosaicism occurs resulting in cells with different genotypes

Engineering Contradiction:

Improvegene editing precisionVSAvoidgenetic uniformity

Core Design Contradiction:

Manufacturing precisionVSStability of the object's composition

Solution Approach 1:

The invention applies CRISPR-based gene editing at the earliest possible stage - the zygote or one-cell embryo stage - before the first cell division occurs. By performing the genetic correction before the embryo begins to divide into multiple cells, the modification is established in the single-cell state, ensuring that all subsequent cells inherit the same corrected genotype, thereby preventing mosaicism.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The invention utilizes the embryo's own cellular machinery and natural DNA repair mechanisms to complete the correction process. After the RNA-guided endonuclease creates a double-strand break at the targeted location, the embryo's endogenous repair systems automatically repair the break, often using the homologous chromosome as a template, thereby achieving precise correction without requiring external intervention for each repair event.

Inventive Principle:

Principle #25Self-service

3Reliability

If maternal and paternal genomes are physically separated in separate nuclei during the first cell cycle, then genome protection is maintained, but recombination events between homologs are limited

Engineering Contradiction:

Improvegenome integrityVSAvoidrecombination capability

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The invention introduces RNA-guided endonucleases as intermediary molecules that can access both the maternal and paternal genomes even when they are physically separated in different nuclei. These endonucleases are delivered into the cytoplasm and can traverse or access both nuclear compartments, enabling targeted DNA cleavage in either nucleus without requiring the genomes to be physically mixed or in close proximity.

Inventive Principle:

Principle #24Intermediary (Mediator)

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 efficient and precise correction of aneuploidies and frame shift mutations, reducing the burden of inherited disorders and improving fertility treatments by ensuring that corrections are made in a non-mosaic fashion, thereby ensuring all cells in the embryo are affected uniformly.

Implementation Method 1

The use of RNA-guided endonucleases, specifically CRISPR/Cas systems, to introduce targeted double-stranded breaks in embryos

Methodology Applied

Scientific EffectCRISPR/Cas system:

Implementation Method 2

The targeted introduction of a DSB followed by recombination allows for the precise modification of genomes

Methodology Applied

Scientific EffectDouble-strand break:

Implementation Method 3

DSBs occur naturally during meiosis, and are repaired through recombination between homologous chromosomes

Methodology Applied

Scientific EffectHomologous recombination:

Implementation Method 4

The correction of pathogenic mutations through interhomolog recombination with a lack of mosaicism

Methodology Applied

Scientific EffectInterhomolog recombination:

Data Source

PatentUS20220323609A1Gene editing to correct aneuploidies and frame shift mutations

Publication Date: 2022.10.13 THE TRUSTEES OF COLUMBIA UNIV IN THE CITY OF NEW YORK

AI summary

The present disclosure relates to using CRISPR-based methods to perform gene editing to correct frame shift mutations in alleles with detectable phenotypes, and to correct aneuploidies.