Self-Delivering Oligonucleotide Editing for Protein-Free Gene Correction
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Solution Overview
Problem
Current nucleic acid editing technologies face challenges such as low efficiency, requirement for exogenous proteins or nucleic acids, immunogenicity, complexity, and inability to efficiently repair point mutations, small insertions, and deletions, while also causing off-target modifications and requiring delivery vehicles.
Innovation Solution
Utilization of single-stranded oligonucleotides that are complementary to DNA or RNA sequences, with chemical modifications to enhance nuclease stability and cellular uptake, allowing for site-specific editing without the need for exogenous proteins or nucleic acids, and capable of correcting point mutations, insertions, and deletions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If chemical modification mechanism is used for nucleic acid editing, then site-specific chemical correction can be achieved, but editing efficiency remains low and requires exogenous proteins or nucleic acids
Solution Approach 1:
The patent employs a self-delivering oligonucleotide that utilizes the cell's own replication machinery to achieve editing without external protein or nucleic acid assistance. The oligonucleotide binds to the target sequence and leverages cellular enzymes and processes to effect the chemical modification, thereby achieving site-specific editing while eliminating the need for exogenous editing tools.
Solution Approach 2:
The patent introduces chemical modifications to the oligonucleotide structure (such as phosphorothioate backbones, 2'-O-methyl modifications, or other chemically modified nucleotides) to enhance its stability, binding affinity, and catalytic activity. These parameter changes in the oligonucleotide's chemical properties enable more efficient editing while maintaining site-specificity.
2Productivity
If CRISPR/Cas-9 gene editing approach is used, then editing efficiency is enhanced, but immunogenicity and complexity increase
Solution Approach 1:
The patent extracts and eliminates the need for exogenous proteins (such as Cas9) and complex delivery vehicles from the editing system. By using a self-delivering oligonucleotide that exploits cellular processes, the system achieves editing efficiency without the immunogenicity and complexity associated with CRISPR/Cas-9 and other protein-based systems.
Solution Approach 2:
The patent employs a small, chemically modified oligonucleotide as the editing agent, replacing large, complex, and potentially immunogenic protein systems. This small molecule approach is simpler to deliver, less immunogenic, and achieves the desired editing function without the complexity of CRISPR/Cas-9 systems.
3Reliability
If exogenous proteins or nucleic acids are required for editing, then editing can be achieved, but delivery vehicles and immunogenicity issues arise
Solution Approach 1:
The self-delivering oligonucleotide utilizes the cell's inherent replication and repair machinery to perform editing without requiring external protein or nucleic acid delivery systems. This eliminates the need for complex delivery vehicles and reduces immunogenicity by avoiding foreign protein expression.
Solution Approach 2:
The patent replaces mechanical/protein-based delivery systems (such as viral vectors or protein transduction) with a chemical oligonucleotide approach that enters cells through standard endocytic pathways and utilizes cellular enzymes for editing, thereby reducing immunogenicity and delivery complexity.
4Measurement precision
If conventional editing methods are used, then point mutations can be targeted, but small insertions and deletions cannot be efficiently repaired
Solution Approach 1:
The patent designs a universal editing platform using self-delivering oligonucleotides that can efficiently repair various types of genetic lesions including point mutations, small insertions, and deletions. The oligonucleotide's ability to bind to target sequences and leverage cellular repair machinery makes it versatile for correcting different mutation types without requiring separate specialized systems.
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 method achieves efficient and specific editing of genomic sequences, improving health outcomes by correcting genetic disorders and reducing disease progression without causing significant off-target effects.
Implementation Method 1
single-stranded oligonucleotides that are complementary to DNA or RNA sequences
Implementation Method 2
chemical modifications to enhance nuclease stability and cellular uptake, allowing for site-specific editing
Data Source
AI summary
The present invention includes compositions and methods for the treatment of a medical condition or disease utilizing editing oligonucleotides. The editing oligonucleotides contain an oligonucleotide strand of about 10 to about 50 nucleotides on each side of the editing moiety which may contain a sugar or linker that positions the active editing moiety in the proper location for hybridization to the target nucleic acid. The editing oligonucleotides may also contain at least one nucleotide sequence change from the targeted sequence in the genome. The method includes modifying a genomic sequence within a cell utilizing an editing oligonucleotide without additional proteins or nucleic acids to assist in the editing process. The editing oligonucleotide may comprise backbone modifications that increase the nuclease stability of the oligonucleotide as compared to unmodified oligonucleotides or oligonucleotides having three phosphorothioates on each terminus.