Artificial Nucleic Acids for RNA Editing via Endogenous Deaminase Recruitment
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Solution Overview
Problem
Current RNA editing strategies face challenges in recruiting endogenous deaminases efficiently and achieving high specificity, often resulting in off-target editing due to hyperactive mutants like E/Q, which enhance editing efficiency but compromise specificity.
Innovation Solution
Development of artificial nucleic acids with chemically modified targeting sequences and recruiting moieties that specifically recruit endogenous deaminases to target RNA sites, enhancing editing efficiency while minimizing off-target effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If hyperactive mutants like E/Q are used to improve editing efficiency, then productivity increases, but manufacturing precision deteriorates due to off-target editing
Solution Approach 1:
The patent modifies the recruiting moiety sequence parameters to optimize deaminase recruitment while maintaining specificity. By changing the nucleotide sequence composition and structure of the recruiting moiety, the system achieves high editing efficiency without the off-target effects associated with hyperactive mutants like E/Q
Solution Approach 2:
The patent applies different functional properties to different parts of the artificial nucleic acid molecule. The recruiting moiety is specifically designed with sequences that selectively recruit endogenous deaminases to target sites, while the rest of the molecule maintains high specificity. This local optimization allows efficient recruitment without compromising overall specificity
2Device complexity
If conventional RNA editing strategies are used to achieve editing, then simplicity is maintained, but reliability deteriorates due to insufficient recruitment of endogenous deaminases
Solution Approach 1:
The patent introduces a recruiting moiety as an intermediary component that facilitates the interaction between artificial nucleic acids and endogenous deaminases. This recruiting moiety acts as a mediator that enhances recruitment efficiency without requiring complex external systems or ectopic protein expression, thereby maintaining relative simplicity while improving reliability
3Manufacturing precision
If site-directed RNA editing is performed to achieve high specificity, then manufacturing precision improves, but productivity deteriorates due to low editing yields
Solution Approach 1:
The patent optimizes the parameters of the recruiting moiety to enhance deaminase recruitment and catalytic activity at target sites. By modifying the sequence composition, length, and structural properties of the recruiting moiety, the system achieves both high specificity and high editing yields, resolving the trade-off between precision and productivity
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 artificial nucleic acids achieve high yields of specific RNA editing with reduced off-target editing, demonstrating improved stability and applicability across various cell types, including primary human cells, with enhanced serum stability.
Implementation Method 1
The artificial nucleic acid comprises a targeting sequence, which comprises a nucleic acid sequence complementary to a target sequence in a target RNA
Implementation Method 2
Targeting such enzyme activities to specific sites at selected transcripts, a strategy called site-directed RNA editing, holds great promise for the treatment of disease
Data Source
AI summary
The present invention concerns artificial nucleic acids for site-directed editing of a target RNA. In particular, the present invention provides artificial nucleic acids capable of site-directed editing of endogenous transcripts by harnessing an endogenous deaminase. Further, the present invention provides artificial nucleic acids for sited-directed editing of a target RNA, which are chemically modified, in particular according to a modification pattern as described herein. The invention also comprises a vector encoding said artificial nucleic acid and a composition comprising said artificial nucleic acid. Moreover, the invention provides the use of the artificial nucleic acid, the composition or the vector for site-directed editing of a target RNA or for in vitro diagnosis. In addition, the artificial nucleic acid, the composition or the vector as described herein are provided for use as a medicament or for use in diagnosis of a disease or disorder.