Mitochondrial Genome Editing via Enzymatic Delivery

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Solution Overview

Problem

Current therapies are inadequate for treating mitochondrial encephalomyopathies due to the inability to transform mitochondrial DNA, resulting in over 600 known mtDNA mutations with diverse clinical features and no approved clinical treatments.

Innovation Solution

A composition including a delivery vehicle with a protein covalently attached to a mitochondrial localization amino acid sequence and an RNA-guided DNA endonuclease enzyme, such as Cas9 or Cpf1, is introduced to target and alter mitochondrial nucleic acid sequences, allowing for the treatment of mitochondrial disorders.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional therapies are used for mitochondrial encephalomyopathies, then treatment is impossible, but the inability to transform mitochondrial DNA prevents any genetic modification

Engineering Contradiction:

Improvetreatment effectivenessVSAvoidability to transform mitochondrial DNA

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent uses a delivery vehicle (lipid nanoparticle or cell-penetrating peptide) as an intermediary to transport the Cas9 protein and guide RNA into mitochondria. This mediator overcomes the natural barrier that prevents DNA transformation in mitochondria, enabling genetic modification without requiring direct transformation capability

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces the conventional mechanical/transformation-based DNA modification approach with an RNA-guided enzymatic system. Instead of using traditional transformation methods that require cellular uptake mechanisms, the system uses guide RNA to direct Cas9 to specific mitochondrial DNA sequences for precise editing

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Adaptability or versatility

If mitochondrial DNA is targeted for editing, then therapeutic potential is achieved, but delivery to mitochondria is extremely difficult

Engineering Contradiction:

Improvetherapeutic applicabilityVSAvoiddelivery system complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent segments the delivery system into distinct functional components: a delivery vehicle (lipid nanoparticle or cell-penetrating peptide), the Cas9 protein, and guide RNA. This segmentation allows each component to be optimized independently for its specific function while simplifying the overall delivery challenge

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent changes the physical and chemical parameters of the delivery system by using lipid nanoparticles or cell-penetrating peptides that can traverse mitochondrial membranes. These parameter changes enable the system to overcome the impermeability barrier of mitochondrial membranes without complex engineering

Inventive Principle:

Principle #35Parameter changes

3Reliability

If Cas9 protein is delivered to mitochondria, then mitochondrial DNA editing is enabled, but protein localization to mitochondria must be achieved

Engineering Contradiction:

Improvemitochondrial DNA editing capabilityVSAvoidprotein localization efficiency

Core Design Contradiction:

ReliabilityVSEase of operation

Solution Approach 1:

The patent uses a mitochondrial targeting sequence as an intermediary that directs the Cas9 protein to mitochondria. This peptide mediator recognizes and facilitates transport of the Cas9 protein across mitochondrial membranes, solving the localization problem without requiring complex cellular machinery

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent extracts the mitochondrial targeting sequence from its native context and attaches it to the Cas9 protein. This extraction and repositioning of the targeting sequence allows direct control over Cas9 localization to mitochondria, simplifying the operation of achieving protein localization

Inventive Principle:

Principle #2Taking out (Extraction)

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 effective targeting and alteration of mitochondrial DNA, potentially providing a therapeutic solution for mitochondrial disorders by delivering the necessary genetic modifications directly to mitochondria.

Implementation Method 1

a protein including a mitochondrial localization amino acid sequence covalently attached to an RNA-guided DNA endonuclease enzyme

Methodology Applied

Scientific EffectProtein targeting and localization:

Implementation Method 2

an RNA-guided DNA endonuclease enzyme, such as Cas9 or Cpf1, is introduced to target and alter mitochondrial nucleic acid sequences

Methodology Applied

Scientific EffectRNA-DNA hybridization and enzymatic cleavage:

Implementation Method 3

a composition including a delivery vehicle with a protein covalently attached to a mitochondrial localization amino acid sequence

Methodology Applied

Scientific EffectCellular uptake and membrane translocation:

Data Source

PatentUS11779657B2Compositions and methods for mitochondrial genome editing

Publication Date: 2023.10.10 RGT UNIV OF CALIFORNIA

AI summary

Compositions and methods for mitochondria genome editing are provided. Also provided are methods for treating mitochondrial disorders by the disclosed compositions.