Antisense Oligonucleotides for CEP290 Splicing Correction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current gene augmentation therapies, such as those using adeno-associated vectors, are limited in treating Leber congenital amaurosis caused by CEP290 mutations due to cargo size constraints and difficulty in controlling expression levels, especially for genes like CEP290 with larger cDNAs, and existing therapies struggle to effectively address the aberrant splicing caused by intronic mutations.
Innovation Solution
Development of viral vectors expressing exon skipping antisense oligonucleotides that specifically target and exclude the aberrant exon from CEP290 mRNA, using sequences complementary to identified nucleotide sequences, to restore normal splicing and protein production, delivered via AAV vectors to modulate splicing and treat CEP290-related diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If gene augmentation therapy using AAV vectors is used to treat LCA, then therapeutic effect is improved for small genes like RPE65, but treatment becomes ineffective for larger genes like CEP290 due to cargo size constraints
Solution Approach 1:
The invention extracts and removes the problematic intron 26 containing the cryptic splice donor site from the CEP290 pre-mRNA through antisense oligonucleotide-mediated exon skipping. This allows the therapeutic approach to bypass the need to deliver the entire large CEP290 gene, instead targeting only the specific defective region, thereby resolving the cargo size limitation of AAV vectors.
Solution Approach 2:
The invention introduces antisense oligonucleotides as intermediary molecules that bind to the cryptic splice donor site in intron 26, preventing aberrant splicing. These oligonucleotides serve as mediators between the AAV vector delivery system and the CEP290 gene, enabling targeted correction of splicing defects without requiring delivery of the entire large gene sequence.
2Reliability
If gene replacement therapy is used to treat LCA, then functional protein production is improved, but control over expression levels becomes difficult
Solution Approach 1:
Instead of replacing the entire CEP290 gene, the invention applies partial action by targeting only the specific cryptic splice donor site in intron 26 with antisense oligonucleotides. This partial targeting approach restores normal splicing of the endogenous gene while maintaining its native regulation mechanisms, thereby achieving functional protein production with preserved expression control.
Solution Approach 2:
The invention enables the endogenous CEP290 gene to serve itself by using its own regulatory elements and splicing machinery. The antisense oligonucleotides temporarily correct the splicing defect, allowing the gene to resume its normal self-regulated expression without requiring external control mechanisms from a replacement gene.
3Ease of operation
If traditional therapy approaches are used for LCA, then treatment simplicity is maintained, but ability to address aberrant splicing caused by intronic mutations is insufficient
Solution Approach 1:
The invention replaces traditional mechanical gene delivery approaches with a molecular biology-based approach using antisense oligonucleotides. These oligonucleotides specifically bind to and block the cryptic splice donor site through base pairing, providing precise molecular-level correction of splicing defects while maintaining relative treatment simplicity through AAV-mediated delivery.
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 effectively redirects normal splicing of CEP290 mRNA, increasing wild-type protein synthesis, potentially delaying or preventing the progression of Leber congenital amaurosis by specifically targeting and correcting aberrant splicing defects, offering a therapeutic strategy for CEP290-associated conditions.
Implementation Method 1
antisense oligonucleotide that binds to and/or is complementary to a polynucleotide with the nucleotide sequence as shown in SEQ ID NO: 17
Data Source
Figure 1A~1C
Figure 2a
Figure 2b
AI summary
The present 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 Leber congenital amaurosis.