Optimized OPA1 Nucleotide Sequence for ADOA Gene Therapy
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Solution Overview
Problem
Current gene therapy approaches for autosomal dominant optic atrophy (ADOA) face challenges due to low expression amounts and poor therapeutic effects of recombinant human type II mitochondrial dynein-like GTPase, primarily because of inefficient gene sequences and varied OPA1 mutations leading to truncated proteins with reduced GTPase activity.
Innovation Solution
An optimized nucleotide sequence for the human mitochondrial dynein-like GTPase is developed, specifically designed for efficient transcription and expression in mammalian cells, using an adeno-associated virus vector to increase the expression and activity of the OPA1 protein, thereby addressing the pathogenic insufficiency in ADOA.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional gene therapy approaches are used for ADOA, then treatment is attempted, but the expression amount of recombinant OPA1 protein is low and therapeutic effects are poor
Solution Approach 1:
The patent applies parameter changes by optimizing the nucleotide sequence of the OPA1 gene through codon optimization and modifying the 5'UTR and 3'UTR regions. This changes the molecular parameters of the gene to enhance transcription and translation efficiency, resulting in significantly increased OPA1 protein expression levels and improved therapeutic effects in ADOA treatment models.
2Adaptability or versatility
If various forms of OPA1 mutations are present in patients, then diverse pathogenic proteins are produced, but a single gene drug cannot treat all patients
Solution Approach 1:
The patent achieves universality by creating a universal therapeutic nucleotide sequence that can treat multiple types of OPA1 mutations. The optimized sequence is designed to work across different mutation variants, allowing a single gene therapy product to address diverse pathogenic forms of OPA1 deficiency, thereby expanding treatment coverage without requiring separate therapies for each mutation type.
3Reliability
If mutant OPA1 encodes truncated proteins lacking complete GTPase domains, then haplotype insufficiency occurs, but the root cause cannot be fully corrected
Solution Approach 1:
The patent applies preliminary action by providing the complete, optimized OPA1 nucleotide sequence before protein synthesis occurs. This ensures that the full-length, functional OPA1 protein with complete GTPase domains is produced from the outset, preventing the formation of truncated proteins. The optimized sequence includes all necessary coding regions and regulatory elements to ensure proper protein folding and function, thereby restoring mitochondrial haplotype insufficiency at its source.
Data Source
AI summary
A gene sequence of recombinant human type II mitochondrial dynein-like GTPase having a nucleotide sequence shown in SEQ ID NO: 1 and uses thereof. A fusion nucleic acid comprising a nucleic acid encoding human type II mitochondrial dynein-like GTPase. A recombinant expression vector comprising the nucleic acid or a fusion nucleic acid. A transformant by which the nucleic acid or the fusion nucleic acid is introduced into a host. A non-human mammalian ADOA model based on the inactivation of the gene of type II mitochondrial dynein-like GTPase, which can effectively improve the pathological manifestations of ADOA using a recombinant expression vector encoding the human type II mitochondrial dynein-like GTPase. The expression level of the nucleic acid encoding the human type II mitochondrial dynein-like GTPase is higher, therefore, more human type II mitochondrial dynein-like GTPase can be obtained in the mitochondria. which can better treat eye diseases such as ADOA.