Antisense Oligonucleotide Corrects Mutant DDC Gene Splicing
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Solution Overview
Problem
AADC deficiency caused by the IVS6+4A>T mutation leads to aberrant splicing of the dopa decarboxylase gene, resulting in incorrect mRNA processing and reduced serotonin levels, for which current treatments lack effective solutions.
Innovation Solution
Designing specific antisense oligonucleotides (ASOs) that target the IVS6+4A>T mutation in the dopa decarboxylase gene to correct aberrant splicing, using morpholino ASOs for stability and specificity, and delivering them into cells to restore normal splicing patterns and increase serotonin production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If the IVS6+4A>T mutation occurs in the DDC gene, then splicing is adjusted to a cryptic site, but this causes aberrant gene splicing with extra 37 base pairs inserted into intron 6
Solution Approach 1:
The patent introduces antisense oligonucleotides as intermediary molecules that bind to the mutant DDC gene pre-mRNA at the cryptic splice site region. These ASOs act as mediators that block the aberrant splicing mechanism caused by the IVS6+4A>T mutation, preventing the insertion of extra 37 base pairs and restoring normal splicing patterns.
Solution Approach 2:
The patent changes the molecular parameters of the splicing process by introducing exogenous oligonucleotide sequences that alter the binding affinity and splicing efficiency. By modifying the nucleotide sequence parameters at the splice site through ASO binding, the patent restores proper splicing recognition and eliminates the aberrant 37 base pair insertion.
2Reliability
If ASOs are designed to target the IVS6+4A>T mutation, then normal splicing can be restored, but this requires specific sequence design and delivery into cells
Solution Approach 1:
The patent segments the therapeutic approach into distinct components: specifically designed antisense oligonucleotide sequences that target the mutant region, delivery mechanisms into cells, and validation of splicing correction. This segmentation allows for systematic optimization of each component's sequence and delivery parameters to achieve reliable splicing correction.
3Ease of operation
If current treatments are used for AADC deficiency, then symptom management is attempted, but no effective solutions exist for the underlying splicing defect
Solution Approach 1:
The patent converts the harmful effect of the IVS6+4A>T mutation into a beneficial therapeutic target. By designing ASOs that specifically recognize and bind to the mutant sequence, the patent transforms the aberrant splicing defect into a precise target for molecular correction, enabling restoration of normal DDC gene function and serotonin production.
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 ASOs successfully restore normal splicing isoforms, reduce aberrant splicing isoforms, and increase serotonin levels in cells with the IVS6+4A>T mutation, providing a potential therapeutic approach for AADC deficiency.
Implementation Method 1
antisense oligonucleotides (ASOs) is a kind of artificial syntheses single strand DNA or RNA molecular with about 13-45 nucleotides which can hybridize to the complement mRNA sequence
Implementation Method 2
different types of ASO can adjust aberrant gene splicing successfully by delivering it into cells
Data Source
AI summary
This present invention discloses an antisense oligonucleotide for splicing adjustment of mutant dopa decarboxylase gene which is complementary to SEQ ID NO: 1. This antisense oligonucleotide can modulate alternative splicing site of mutant dopa decarboxylase gene. It is helpful to research and develop drug to treat AADC deficiency symptom. This present invention also discloses a method to use said antisense oligonucleotide in vitro.