Double-Stranded Oligonucleotide for ADAR-Mediated RNA Editing
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Solution Overview
Problem
Current synthetic oligonucleotides are not capable of selectively editing target RNAs in a therapeutically effective manner using ADAR proteins, which are essential for converting adenosine to inosine in specific mRNAs, leading to potential changes in protein function.
Innovation Solution
A double-stranded oligonucleotide comprising a guide oligonucleotide and a passenger oligonucleotide, where the guide oligonucleotide is at least 70% to 90% complementary to the passenger oligonucleotide, and the second portion is complementary to a target mRNA, facilitating ADAR-mediated adenosine to inosine conversion at specific sites in target mRNAs such as SERPINA1, LRRK2, and others.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If synthetic oligonucleotides are used to edit target RNAs via ADAR proteins, then adenosine deamination can occur, but the editing is not therapeutically effective or selective enough
Solution Approach 1:
The oligonucleotide is divided into three functional segments: a 5' portion (70-90% complementary to passenger strand for ADAR recruitment), a middle portion (completely complementary to target mRNA for precise positioning), and a 3' portion (70-90% complementary to passenger strand for stable duplex formation). This segmentation allows each region to perform its specific function optimally, achieving both therapeutic effectiveness and editing selectivity simultaneously.
2Reliability
If the guide oligonucleotide is highly complementary to the passenger oligonucleotide, then ADAR recruitment is enhanced, but specificity to the target mRNA may be reduced
Solution Approach 1:
Different regions of the guide oligonucleotide have different complementarity qualities: the 5' and 3' portions have 70-90% complementarity to promote ADAR binding and duplex stability, while the middle portion has 100% complementarity to the target mRNA for precise positioning. This local variation in quality allows simultaneous optimization of ADAR recruitment and target specificity.
3Ease of manufacture
If the oligonucleotide structure is simplified, then ease of manufacture increases, but the ability to achieve therapeutically effective editing decreases
Solution Approach 1:
The invention optimizes specific parameters of the oligonucleotide structure: the guide strand is designed at 21-25 nucleotides with 70-90% complementarity to the passenger strand, and the middle portion is precisely 7-15 nucleotides long. These parameter optimizations enable effective ADAR-mediated editing while maintaining synthetic feasibility through standard oligonucleotide synthesis methods.
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 described oligonucleotide effectively recruits ADAR enzymes to specific adenosine residues in target mRNAs, enabling precise editing and potential therapeutic interventions for diseases associated with single nucleotide polymorphisms by altering codon sequences and protein function.
Implementation Method 1
ADARs are enzymes that bind to double-stranded RNA (dsRNA) and convert adenosine to inosine through deamination
Implementation Method 2
the guide oligonucleotide comprises a first portion and a second portion, wherein the first portion is at least 70%, at least 80%, at least 85%, or at least 90% complementary to the passenger oligonucleotide, and wherein the second portion is complementary to a target mRNA
Data Source
AI summary
The present invention relates to methods and compositions for editing a polynucleotide. e.g., a polynucleotide comprising a SNP associated with a disease or disorder.