Allele-Specific CRISPR Knockout for Retinitis Pigmentosa

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods are inadequate for distinguishing and knocking out dominant-mutated alleles causing genetic disorders like retinitis pigmentosa, as they fail to specifically target and inactivate the mutated proteins while allowing expression of functional proteins.

Innovation Solution

A method utilizing CRISPR nuclease and RNA molecules with guide sequences to specifically target and degrade the mRNA of dominant-mutated alleles, allowing expression of functional alleles, thereby treating or preventing genetic disorders such as retinitis pigmentosa.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If current methods are used to target mutated alleles, then general gene targeting is achieved, but specific distinction between mutated and functional alleles is lost

Engineering Contradiction:

Improveallele discrimination precisionVSAvoidmethod complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The guide RNA is designed to target a specific local region of the mutated allele that contains a unique nucleotide difference from the functional allele. This localized targeting approach enables precise discrimination between alleles without requiring complex global identification methods, thereby improving measurement precision while maintaining relatively simple device complexity.

Inventive Principle:

Principle #3Local quality

2Reliability

If dominant-mutated alleles are knocked out, then disease phenotype is reduced, but expression of functional proteins must be preserved

Engineering Contradiction:

Improvedisease treatment efficacyVSAvoidloss of functional protein

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The method segments the Rhodopsin gene into two distinct alleles: the mutated allele and the functional allele. By designing the CRISPR-Cas9 system with a guide RNA that specifically targets only the mutated allele through unique nucleotide differences, the system can selectively knock out the harmful mutated allele while leaving the functional allele intact and expressed, thereby improving disease treatment efficacy without causing loss of functional protein.

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 effectively inactivates the mutated alleles, reducing the disease phenotype by allowing the expression of functional proteins, thereby ameliorating or preventing genetic disorders like retinitis pigmentosa.

Implementation Method 1

A method utilizing CRISPR nuclease and RNA molecules with guide sequences to specifically target and degrade the mRNA of dominant-mutated alleles

Methodology Applied

Scientific EffectCRISPR-Cas9 RNA-guided DNA/RNA cleavage:

Implementation Method 2

an RNA molecule comprising a guide sequence portion having 17-20 nucleotides in the sequence of 17-20 contiguous nucleotides set forth in any one of SEQ ID NOs: 1-3010

Methodology Applied

Scientific EffectNucleic acid base pairing (hybridization):

Data Source

PatentUS20230173105A1Differential knockout of an allele of a heterozygous rhodopsin gene

Publication Date: 2023.06.08 EMENDOBIO INC

AI summary

RNA molecules comprising a guide sequence portion having 17-20 nucleotides in the sequence of 17-20 contiguous nucleotides set forth in any one of SEQ ID NOs: 1-3010 and compositions, methods, and uses thereof