USH2A Exon 68 Antisense Oligonucleotides for Exon Skipping
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current treatments for Usher syndrome associated with USH2A mutations are limited, particularly due to the large size of the usherin-encoding sequence exceeding the packaging capacity of conventional gene delivery vectors, and there is a need for effective therapies to address mutations in regions other than exon 13.
Innovation Solution
Development of antisense oligonucleotides (ASOs) that target and skip exon 68 of the USH2A gene, which are designed to bind specifically to the exon sequence and are delivered via viral vectors like AAV, to modulate splicing and restore usherin protein function.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional gene augmentation therapy using AAV or LV vectors is used, then the packaging capacity is limited to 4.7 kb or 8 kb respectively, but the usherin-encoding sequence is 15.6 kb which exceeds this capacity
Solution Approach 1:
The invention extracts and removes specific exons (exon 68 or exon 71) from the usherin pre-mRNA transcript using antisense oligonucleotides. This exon skipping approach allows the final protein product to be generated from a reduced transcript that can be accommodated within viral vector packaging limits, while still producing a functional usherin protein that rescues retinal phenotype.
Solution Approach 2:
The invention changes the splicing parameters of the USH2A gene by using antisense oligonucleotides to modulate pre-mRNA splicing. This results in exon skipping and produces a shortened but functional usherin protein isoform that can be effectively delivered and expressed within the constraints of AAV vector packaging capacity.
2Adaptability or versatility
If ASO-induced exon skipping is used, then the packaging capacity constraint is overcome, but the therapy must be mutation-independent to address the diversity of USH2A mutations
Solution Approach 1:
The invention develops a universal ASO-based therapy that can address multiple different USH2A mutations through a common mechanism. The antisense oligonucleotides target conserved splicing regulatory elements that are present across different mutant alleles, allowing a single therapeutic approach to be effective for patients with diverse loss-of-function mutations in the USH2A gene.
3Reliability
If exon 13 skipping is performed, then clinical benefit is achieved in multiple visual function parameters, but this approach only addresses mutations in or near exon 13
Solution Approach 1:
The invention segments the USH2A gene into different targetable exons (exon 68 and exon 71) that can be individually skipped using ASOs. This segmentation allows the development of multiple ASO candidates targeting different regions of the gene, thereby expanding the coverage to address mutations distributed throughout the 72-exon USH2A transcript.
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 ASO-induced skipping of exon 68 restores usherin protein expression and rescues rhodopsin localization to the photoreceptor outer segment, providing a promising treatment for USH2A-associated retinitis pigmentosa.
Implementation Method 1
antisense oligonucleotide for skipping of exon 68 that binds to and/or is complementary to a polynucleotide with the nucleotide sequence as shown in SEQ ID NO: 1
Data Source
AI summary
The invention relates to the fields of medicine and immunology. In particular, it relates to novel antisense oligonucleotides that may be used in the treatment, prevention and/or delay of an USH2A-related disease or condition.