Trans-Splicing RNA Tethering for Efficient Exon Replacement
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Solution Overview
Problem
Current methods for treating human genetic diseases are inadequate in efficiently replacing defective genetic sequences in human cells.
Innovation Solution
A system for trans-splicing using a nucleic acid molecule encoding an exonic sequence and intronic domain, along with an RNA-binding protein, to insert the exonic sequence into a target RNA molecule, without relying on CRISPR-associated proteins, utilizing tethering proteins and engineered small nuclear RNAs to promote efficient insertion and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CRISPR-associated proteins are used for genetic sequence replacement, then insertion capability is improved, but system complexity and potential off-target effects increase
Solution Approach 1:
The patent extracts and removes the CRISPR-associated protein component from the system, replacing it with a simplified trans-splicing mechanism that uses intronic domains and RNA-binding proteins to achieve genetic sequence replacement without the complexity and off-target effects associated with CRISPR systems
Solution Approach 2:
The patent introduces intronic domains as intermediary elements that mediate the insertion process. These intronic domains contain binding sites for RNA-binding proteins and facilitate the trans-splicing reaction, serving as a bridge between the guide RNA and the target genomic sequence without requiring CRISPR-associated proteins
2Productivity
If trans-splicing efficiency is increased through multiple components, then productivity is improved, but device complexity increases
Solution Approach 1:
The patent merges multiple functional elements into a unified trans-splicing system. The intronic domain integrates guide sequence complementarity, binding sites for RNA-binding proteins, and trans-splicing catalytic elements into a single functional unit that operates efficiently without requiring separate complex components
Solution Approach 2:
The patent designs the intronic domain to perform multiple functions simultaneously: it provides guide sequence complementarity for target recognition, contains binding sites for RNA-binding proteins to facilitate recruitment, and catalyzes the trans-splicing reaction. This multi-functionality increases efficiency while maintaining system simplicity
Data Source
AI summary
Described herein are compositions systems and methods for promoting trans-splicing. In some examples, the system may comprise a nucleic acid molecule. The nucleic acid molecule may encode an exonic sequence. The system may further comprise a tethering fusion protein. The tethering fusion protein may promote an association of the exonic sequence and a target RNA or portion thereof.