ED-RgE Fusion Protein for Targeted Splicing Modulation
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Solution Overview
Problem
Current gene therapeutic approaches lack effective methods for modulating the splicing process, particularly for targeted rescue or induction of missplicing events, which are crucial for treating inherited retinal diseases.
Innovation Solution
A gene editing system comprising an RNA-guided endonuclease molecule fused with exonuclease activity, paired with a single guide RNA, is designed to associate with genomic DNA at splicing sites, causing deletions that modulate splicing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current gene therapeutic approaches are used, then treatment of inherited retinal diseases can be attempted, but effective modulation of splicing process is lacking
Solution Approach 1:
The patent combines RNA-guided endonuclease activity with exonuclease activity into a single fusion protein (ED-RgE). This merging of two distinct enzymatic functions enables the system to both target specific splicing sites through RNA guidance and efficiently modulate splicing through exonuclease-mediated DNA degradation, thereby achieving effective splicing modulation while maintaining versatility in sgRNA design
Solution Approach 2:
The ED-RgE fusion protein serves multiple functions: it acts as an RNA-guided endonuclease for targeted DNA cleavage, an exonuclease for DNA degradation and modulation, and works with versatile sgRNA molecules that can target various splicing sites. This multi-functionality allows the system to address different splicing defects across various inherited retinal diseases, enhancing both reliability and adaptability
2Adaptability or versatility
If targeted splicing modulation is achieved, then therapeutic options for genetic diseases are expanded, but the system complexity increases
Solution Approach 1:
By fusing the RNA-guided endonuclease and exonuclease into a single ED-RgE fusion protein, the patent reduces system complexity compared to using separate proteins. This merged architecture simplifies delivery and expression while maintaining the ability to perform targeted splicing modulation, thereby expanding therapeutic options without proportionally increasing system complexity
Solution Approach 2:
The universal ED-RgE fusion protein can be paired with different sgRNA molecules to target various splicing sites in different genes associated with inherited retinal diseases. This universal platform approach allows diverse therapeutic applications using a single core system, expanding adaptability while keeping the base system relatively simple
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 system enables targeted modulation of splicing, achieving higher splicing rescue efficiency and flexibility in sgRNA design, thereby providing new therapeutic options for genetic diseases, including inherited retinal diseases.
Implementation Method 1
a nucleic acid encoding an RNA-guided endonuclease molecule or fragment thereof fused to a molecule comprising exonuclease activity (ED-RgE molecule)
Implementation Method 2
fused to a molecule comprising exonuclease activity (ED-RgE molecule)
Implementation Method 3
one single guide RNA (sgRNA) molecule, wherein said ED-RgE molecule and said sgRNA molecule are configured to associate with genomic DNA comprising a splicing site
Data Source
AI summary
The invention is directed to a gene editing system for editing genomic DNA to modulate splicing, a polynucleotide or vector encoding said gene editing system, a lipid particle comprising said gene editing system, a pharmaceutical composition comprising said gene editing system polynucleotide or vector or lipid particle, methods of editing genomic DNA in a cell to modulate splicing, and a cell processed by said methods.