CRISPR-Cas9 Gene Editing for Retinal Dysfunction
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Solution Overview
Problem
Current therapeutic agents for Leber congenital amaurosis (LCA), caused by genetic mutations in over 20 different genes, have limited effectiveness as they only function while the AAV vector is present, necessitating a method for longer-term correction of mutations in retinal function-forming genes like RPE65.
Innovation Solution
A composition for gene manipulation using a guide nucleic acid capable of targeting retinal function-forming genes, specifically the RPE65 gene, combined with editor proteins such as Cas9, to artificially manipulate and correct mutant sequences, thereby treating retinal dysfunction diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If AAV-based therapeutic agents are used to express RPE65, then therapeutic effect is achieved, but the effect is limited to the duration AAV is present
Solution Approach 1:
The therapeutic approach is segmented into two components: (1) AAV vector delivers CRISPR-Cas9 system for gene editing, and (2) guide RNA directs the system to specifically target and correct the RPE65 mutation. This segmentation allows the AAV to serve only as a delivery vehicle rather than a permanent therapeutic agent, enabling long-term correction through one-time genetic modification.
Solution Approach 2:
The CRISPR-Cas9 system performs preliminary action by creating double-strand breaks at the mutant RPE65 gene locus before the AAV vector is cleared from the system. This preliminary gene editing action establishes permanent genetic correction that persists independently of the AAV vector's presence, resolving the duration limitation.
2Duration of action of moving object
If gene manipulation is performed to correct mutant sequences, then long-term therapeutic effect is achieved, but complexity of the treatment increases
Solution Approach 1:
The AAV vector is designed with multi-functionality: it simultaneously delivers both the Cas9 enzyme and the guide RNA sequence in a single vector system. This universal delivery approach simplifies the treatment protocol compared to multiple separate administrations, reducing operational complexity while enabling long-term therapeutic effects through gene correction.
Solution Approach 2:
The guide RNA acts as an intermediary that bridges the Cas9 enzyme and the target RPE65 gene sequence. It provides specific directional guidance to the mutant locus, enabling precise gene manipulation without requiring complex targeting mechanisms, thus simplifying the overall treatment complexity while achieving durable correction.
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
The approach allows for long-term correction of retinal function-forming genes, potentially treating LCA and other retinal dysfunction diseases by ensuring the retinal genes function normally, even after the initial treatment vector is gone.
Implementation Method 1
The guide domain may include a nucleotide sequence capable of forming a complementary binding with a guide nucleic acid-binding sequence of target sequence of the retinal function-forming gene.
Implementation Method 2
a composition for gene manipulation which may include a guide nucleic acid capable of targeting a retinal function-forming gene
Data Source
AI summary
The present invention relates to a composition for gene manipulation for treating or improving a retinal dysfunction disease or a method using the same. More particularly, the present invention relates to a composition for gene manipulation including a guide nucleic acid capable of targeting a retinal function-forming gene and a method of treating or improving a disease caused by retinal dysfunction by artificially manipulating and/or correcting a retinal function-forming gene using the same.