CRISPR-Cas9 NRF2 Knockout for Cancer Chemotherapy Sensitization

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Solution Overview

Problem

Current cancer treatments, particularly chemotherapy, often result in severe side effects and can become ineffective over time due to resistance from cancer cells, highlighting the need for improved methods to treat cancer.

Innovation Solution

The method involves introducing guide RNAs complementary to the NRF2 gene and a CRISPR-associated endonuclease into cells to reduce NRF2 expression or activity, thereby sensitizing cancer cells to chemotherapy and inhibiting tumor growth.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If chemotherapy is used to treat cancer, then cancer cell proliferation is inhibited, but severe side effects occur and cancer cells develop resistance

Engineering Contradiction:

Improvecancer treatment effectivenessVSAvoidside effects and drug resistance

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and targets the specific NRF2 protein responsible for cancer cell resistance and survival. By using CRISPR/Cas9 to deliver guide RNAs that specifically bind to NRF2 mRNA, the system selectively removes or reduces NRF2 expression in cancer cells, thereby eliminating the source of drug resistance without affecting other cellular processes that would cause side effects

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention applies local quality by specifically targeting NRF2 expression in cancer cells while leaving normal cells unaffected. The guide RNAs are designed to recognize and bind to NRF2 mRNA sequences that are specifically upregulated in cancer cells, enabling selective reduction of NRF2 in the tumor microenvironment while preserving normal cellular function in healthy tissues

Inventive Principle:

Principle #3Local quality

2Productivity

If traditional chemotherapy agents are administered, then tumor growth is suppressed, but the treatment becomes ineffective over time due to resistance

Engineering Contradiction:

Improvetumor growth suppressionVSAvoidtreatment effectiveness over time

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention performs preliminary action by reducing NRF2 expression in cancer cells before chemotherapy administration. By using CRISPR/Cas9 to knockdown NRF2 in advance, the cancer cells are rendered more susceptible to chemotherapeutic agents, preventing the development of resistance during subsequent chemotherapy treatment

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The invention changes the key parameter of NRF2 expression levels in cancer cells. By reducing NRF2 mRNA and protein levels through guide RNA-mediated degradation, the system alters the cellular state from a resistant phenotype to a sensitive phenotype, thereby restoring chemotherapy effectiveness

Inventive Principle:

Principle #35Parameter changes

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 decreases cancer cell proliferation and enhances the effectiveness of chemotherapeutic agents like cisplatin and vinorelbine, both in vitro and in a xenograft mouse model, while minimizing side effects.

Implementation Method 1

the one or more gRNAs hybridize to the NRF2 gene and the CRISPR-associated endonuclease cleaves the NRF2 gene

Methodology Applied

Scientific EffectHybridization:

Implementation Method 2

introducing into the cell (a) one or more DNA sequences encoding one or more guide RNAs (gRNAs) that are complementary to one or more target sequences in the NRF2 gene and (b) a nucleic acid sequence encoding a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease

Methodology Applied

Scientific EffectCRISPR/Cas9 gene editing:

Data Source

PatentUS12203070B2Gene knockout of NRF2 for treatment of cancer

Publication Date: 2025.01.21 CHRISTIANA CARE GENE EDITING INSTITUTE INC

AI summary

The disclosure provides a guide RNA (gRNA) comprising a DNA-binding domain and a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease protein-binding domain, wherein the DNA-binding domain is complementary to a target domain from an NRF2 gene. The disclosure also provides nucleic acid sequence encoding the gRNA. The disclosure further provides a method of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease and a guide RNA that is complementary to a target domain from an NRF2 gene in the subject. Methods of treating cancer comprising administering a pharmaceutical composition comprising: a DNA sequence encoding a guide RNA that is complementary to a target domain from an NRF2 gene in the subject; and a nucleic acid sequence encoding a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease, are also provided.