ADAR Guide Oligonucleotides for Precise NRF2-KEAP1 Disruption

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Solution Overview

Problem

Existing methods for disrupting the KEAP1-NRF2 protein interaction lack specificity and stability, making them unsuitable for therapeutic applications.

Innovation Solution

Utilizing guide oligonucleotides to recruit endogenous ADAR enzymes for site-specific adenosine to inosine alterations in polynucleotides encoding NRF2 and KEAP1 proteins, generating mutant amino acids that disrupt their interaction.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If previously disclosed methods using synthetic single-stranded oligonucleotides are used to edit target RNAs, then adenosine deamination can occur, but the methods lack the required specificity, selectivity and stability for therapeutic applications

Engineering Contradiction:

Improvespecificity of RNA editingVSAvoidstability of therapeutic effect

Core Design Contradiction:

Manufacturing precisionVSReliability

Solution Approach 1:

The patent introduces a double-stranded RNA structure as an intermediary that recruits ADAR enzymes to the target site. The dsRNA acts as a mediator between the guide sequence and the adenosine residue, enabling precise and stable editing by bringing the editing enzyme to the correct location through complementary base pairing and secondary structure formation

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the structural parameters of the oligonucleotide from single-stranded to double-stranded RNA, creating stem-loop structures with specific secondary configurations. This parameter change increases the stability and specificity of the editing complex by providing a more rigid and defined structure that resists degradation and maintains proper orientation for ADAR recruitment

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If endogenous ADAR enzymes are recruited for site-specific editing, then precise disruption of NRF2-KEAP1 interaction is achieved, but the complexity of delivering and maintaining editing activity increases

Engineering Contradiction:

Improveprecision of protein interaction disruptionVSAvoidcomplexity of oligonucleotide delivery system

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The dsRNA oligonucleotides are designed to self-recruit endogenous ADAR enzymes through their intrinsic dsRNA structure. The system serves itself by using the natural affinity of ADAR enzymes for dsRNA structures, eliminating the need for external enzyme delivery or complex delivery mechanisms while maintaining precise editing at the target site

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

Instead of delivering engineered editing enzymes to the target site, the patent inverts the approach by delivering a structural element (dsRNA) that naturally attracts the endogenous editing enzymes. This inversion simplifies the delivery system by using the cell's own machinery while achieving the same therapeutic effect

Inventive Principle:

Principle #13The other way round (Inversion)

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 disrupts the NRF2-KEAP1 interaction, increasing expression of protective genes and treating associated diseases without immune response, as it leverages endogenous ADAR enzymes for precise editing.

Implementation Method 1

Adenosine deaminases acting on RNA (ADAR) are enzymes which bind to double-stranded RNA (dsRNA) and convert adenosine to inosine through deamination

Methodology Applied

Scientific EffectDeamination: Hydrolysis

Data Source

PatentUS20250352667A1Methods and compositions for disrupting NRF2-KEAP1 protein interaction by ADAR mediated RNA editing

Publication Date: 2025.11.20 KORRO BIO INC

AI summary

The present invention relates to methods and compositions for disrupting interaction of an NRF2 protein and a KEAP1 protein. The methods include contacting at least one polynucleotide selected from the group consisting of a polynucleotide encoding the NRF2 protein and a polynucleotide encoding the KEAP1 protein with a guide oligonucleotide that effects one or more (e.g., at least two) adenosine deaminase acting on RNA (ADAR)-mediated adenosine to inosine alterations in said at least one polynucleotide, wherein the adenosine to inosine alterations generate a mutant amino acid, thereby disrupting interaction of the NRF2 protein and the KEAP1 protein. The invention also relates to methods of treating a KEAP1-NRF2 pathway related disease in a subject in need thereof, the method comprising contacting, within the subject, at least one polynucleotide selected from the group consisting of a polynucleotide encoding an NRF2 protein and a polynucleotide encoding a KEAP1 protein with a guide oligonucleotide that effects an adenosine deaminase acting on RNA (ADAR)-mediated adenosine to inosine alteration in said at least one polynucleotide, wherein the adenosine to inosine alteration generates a mutant amino acid, thereby disrupting interaction of the NRF2 protein and the KEAP1 protein and treating the disease in the subject; and compositions thereof.