Codon-Optimized RPGR ORF15 Sequence for Stable Gene Therapy
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Solution Overview
Problem
Current gene therapy approaches for X-linked retinitis pigmentosa (XLRP) face challenges due to the mutagenic nature of the RPGR gene, leading to issues with vector stability and functionality, and there is a lack of effective treatments for preventing or improving vision in affected individuals.
Innovation Solution
The development of a codon-optimized RPGR ORF15 sequence linked to a rhodopsin kinase promoter and a bovine growth hormone polyadenylation signal, delivered via an AAV8 vector, which enhances sequence stability and protein expression, reducing mutation occurrence and improving vector fidelity and efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the RPGR gene is used for gene therapy, then treatment of X-linked retinitis pigmentosa is achieved, but mutations occur during vector production leading to reduced vector stability and functionality
Solution Approach 1:
The patent applies codon optimization to the RPGR ORF15 cDNA sequence, modifying nucleotide composition and codon usage patterns while preserving the amino acid sequence. This parameter change in the genetic code reduces mutagenic regions and improves vector production stability without altering the therapeutic protein function.
Solution Approach 2:
The patent creates an optimized copy of the RPGR ORF15 gene sequence that replicates the therapeutic function while eliminating mutagenic properties. This synthetic copy serves as a stable template for vector production, preventing mutation generation during cloning and manufacturing processes.
2Productivity
If gene replacement therapy is developed for XLRP, then vision improvement is achieved, but previous programmes were delayed due to mutations in the transgene cassette
Solution Approach 1:
The patent modifies the nucleotide sequence parameters of RPGR ORF15 through codon optimization, changing GC content and eliminating repetitive sequences that cause instability. This enables reliable large-scale vector production without delays, accelerating therapy development while maintaining genetic integrity.
Solution Approach 2:
The patent identifies and eliminates the mutagenic properties of the original RPGR sequence, converting a harmful characteristic into a beneficial stable sequence. The optimized version maintains therapeutic function while preventing the mutations that previously delayed development programmes.
3Adaptability or versatility
If the RPGR gene is highly mutagenic, then alternative splice variants are generated, but this reduces manufacturing precision of the therapeutic vector
Solution Approach 1:
The patent optimizes codon usage and nucleotide composition in the RPGR ORF15 sequence, changing parameters that affect splicing and mutation. This produces a single, precise therapeutic sequence that maintains protein functionality while eliminating alternative splice variants and improving manufacturing consistency.
Data Source
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AI summary
A polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGRORF15), wherein the RPGRORF15-encoding nucleotide sequence has been codon optimised to increase fidelity of replication of the sequence.