CRISPR-GNDM System Suppresses KRAS Expression
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Solution Overview
Problem
Current therapeutic agents are ineffective in targeting and suppressing KRAS gene expression, particularly in cancers with KRAS mutations, such as pancreatic, lung, and colorectal cancers, where existing drugs like cetuximab and panitumumab fail due to KRAS activating mutations.
Innovation Solution
A CRISPR-Guide Nucleotide Directed Modulation (GNDM) system is employed to suppress KRAS expression by targeting the expression regulatory region of the KRAS gene using dCas9 or dCpf1 proteins, along with guide nucleotides, to recruit transcription repressors like KRAB, thereby reducing KRAS gene expression without inducing double-stranded DNA breaks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional therapeutic agents (cetuximab, panitumumab) are used to target cancer cells, then treatment is effective for cancers without KRAS mutations, but treatment fails in cancers with KRAS activating mutations
Solution Approach 1:
The invention changes the therapeutic approach from targeting downstream effectors (EGFR pathway) to directly targeting the KRAS gene itself using CRISPR-Cas9 genome editing technology. This parameter change in the mechanism of action enables treatment of KRAS-mutant cancers that were previously resistant to conventional therapies.
Solution Approach 2:
The invention introduces CRISPR-Cas9 system as an intermediary mechanism to achieve gene suppression. The guide RNA and Cas9 protein act as intermediaries to specifically target and suppress KRAS gene expression, bridging the gap between conventional therapy limitations and effective treatment of KRAS-mutant cancers.
2Productivity
If low-molecular drug discovery approach is used to find KRAS inhibitors, then extensive screening can be performed, but no effective therapeutic agent has been discovered yet
Solution Approach 1:
The invention replaces the mechanical/chemical approach of low-molecular drug discovery with a biological approach using CRISPR-Cas9 genome editing. Instead of screening thousands of chemical compounds, the system directly edits the KRAS gene to achieve therapeutic effect, fundamentally changing the discovery mechanism from chemical screening to genetic manipulation.
Solution Approach 2:
The invention changes the fundamental parameter of therapeutic intervention from small molecule inhibition to genome editing. This parameter change enables direct targeting of KRAS gene function rather than attempting to bind and inhibit the protein product, overcoming the limitations of conventional drug discovery approaches.
3Reliability
If CRISPR-GNDM system is used to suppress KRAS expression by targeting expression regulatory region, then effective suppression of KRAS gene expression is achieved, but this approach is novel and unproven compared to conventional therapies
Solution Approach 1:
The invention uses CRISPR-Cas9 system as an intermediary to achieve precise gene suppression. The guide RNA and Cas9 protein serve as intermediaries that can be delivered to target cells to suppress KRAS expression, providing a controllable and specific mechanism that overcomes the limitations of conventional therapies while managing system complexity through modular design.
Data Source
AI summary
The present invention provides a method of treating a disease associated with elevated KRAS activity or expression in a subject, comprising suppressing KRAS expression in the subject by targeting an expression regulatory region of KRAS gene using a CRISPR-Guide Nucleotide Directed Modulation (GNDM). Also, provided is a CRISPR-GNDM system for suppressing KRAS expression comprising (a) a protein selected from the group consisting of dCas9 or dCpf1, a fusion protein of dCas9 or dCpf1 and Kruppel associated box (KRAB), and (b) a guide nucleotide targeting an expression regulatory region of KRAS gene.