Antisense Oligonucleotide Exon Skipping for Protease Specificity
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current protease inhibitors have non-specific effects in the human body, leading to unintended consequences and challenges in targeting specific proteases, as they often inhibit multiple proteases, resulting in a range of effects that may not be beneficial for treating diseases effectively.
Innovation Solution
The method involves using antisense oligonucleotides (AONs) to induce skipping of the exon sequence encoding the proteolytic cleavage site in pre-mRNA, preventing its inclusion in the mature mRNA, thereby producing a protein without the cleavage site, which can reduce toxic properties while maintaining desired functions by modifying the splicing process rather than directly inhibiting proteases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If protease inhibitors are used to inhibit proteases, then proteolytic processing is inhibited, but non-specific effects occur and multiple proteases are inhibited
Solution Approach 1:
The invention segments the approach to protease inhibition by targeting the specific mRNA transcript encoding the substrate protein rather than inhibiting the protease enzyme itself. This allows selective removal of the cleavage site from the protein product without affecting other proteases or their substrates in the system
Solution Approach 2:
The invention extracts and removes the specific exon containing the proteolytic cleavage site from the pre-mRNA through antisense-mediated exon skipping. This extracts the harmful element (cleavage site) from the protein coding sequence while preserving the rest of the protein structure and function
2Reliability
If protease inhibitors are designed to be sufficiently specific for the target protease, then specificity is improved, but it becomes difficult to produce such inhibitors
Solution Approach 1:
The invention introduces an intermediary approach by using antisense oligonucleotides to modify mRNA processing rather than directly interacting with the protease enzyme. This intermediary method (targeting RNA splicing) provides a more tractable path to achieving specificity compared to direct protease inhibition
Solution Approach 2:
The invention substitutes the mechanical/enzymatic approach of protease inhibition with a genetic/RNA-based approach. Instead of using small molecule inhibitors or peptides that must physically bind to the protease active site, the invention uses antisense oligonucleotides to modify the genetic blueprint of the substrate protein
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
This approach allows for the selective removal of proteolytic cleavage sites from proteins, reducing toxicity while preserving functional aspects, and is reversible and adjustable, making it suitable for treating diseases associated with proteolytic processing, such as neurodegenerative disorders.
Implementation Method 1
antisense oligonucleotides (AONs) to induce skipping of the exon sequence encoding the proteolytic cleavage site in pre-mRNA
Data Source
AI summary
The invention relates to means and methods for removing a proteolytic cleavage site from a protein comprising providing a cell that expresses pre-mRNA encoding the protein with an anti-sense oligonucleotide that induces skipping of the exonic sequence that encodes the proteolytic cleavage site, the method further comprising allowing translation of mRNA produced from the pre-mRNA.