HbW Polynucleotide Editing for Break-Free Gene Modification
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Solution Overview
Problem
Existing gene editing technologies rely on introducing breaks into target sites for modification, which can be inefficient and may cause off-target effects, particularly in non-replicating cells, and non-human components can trigger immunogenic responses.
Innovation Solution
Development of polynucleotide modification agents comprising a helicase beta-wing element (HbW) and sequence-specific binding elements, which allow for precise genetic modifications without site breaks, applicable to diverse cell types, including primary human cells, and are composed entirely of human sequences to avoid immunogenicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing gene editing technologies introduce breaks into target sites for modification, then genetic modification can be achieved, but off-target effects increase and efficiency decreases in non-replicating cells
Solution Approach 1:
The patent uses a polynucleotide modification agent as an intermediary that binds to the target site and facilitates modification without requiring DNA breaks. The agent comprises a sequence-specific binding element that targets the locus and a helicase beta-wing element that enables modification through a break-free mechanism, thereby achieving reliable editing with reduced off-target effects.
Solution Approach 2:
The patent replaces the mechanical approach of introducing DNA breaks (used in CRISPR-Cas9 and other conventional systems) with a chemical/biological mechanism involving polynucleotide modification agents that modify DNA through alternative pathways such as base editing or epigenetic modification, eliminating the need for double-strand breaks and associated harmful effects.
2Productivity
If non-human components are used in gene modification technologies, then functional gene editing is achieved, but immunogenic responses occur in subjects
Solution Approach 1:
The patent employs human sequences for the polynucleotide modification agent to ensure homogeneity with the host genome. The sequence-specific binding element and helicase beta-wing element are both derived from human genes, eliminating foreign protein expression and subsequent immunogenic responses while maintaining full gene editing functionality in human cells.
3Productivity
If polynucleotide replication is relied upon for modification, then genetic changes can be introduced, but modification fails in non-replicating cells
Solution Approach 1:
The patent segments the gene editing function into two independent components: a sequence-specific binding element that targets the genomic locus and a helicase beta-wing element that performs the modification. This segmentation allows the modification mechanism to operate independently of cell replication, enabling the system to function in both replicating and non-replicating cells.
Solution Approach 2:
The patent changes the fundamental parameter of how modification is achieved from replication-dependent to replication-independent. By using polynucleotide modification agents that can directly modify DNA sequences through mechanisms such as base conversion or methylation rather than relying on replication to fix changes, the system becomes applicable to all cell types including primary human cells that do not divide.
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 agents achieve accurate genetic modifications with low off-target insertions and deletions, effective in both cultured and in vivo conditions, and can be delivered via various modalities, including plasmids, mRNA, and proteins, suitable for non-replicating cells.
Implementation Method 1
a helicase beta-wing element (HbW element) and a sequence-specific binding element
Implementation Method 2
a helicase beta-wing element (HbW element) and a sequence-specific binding element
Data Source
AI summary
The present disclosure provides technologies for genetic modification that use a helicase beta-wing element (HbW element). Provided technologies enable genetic modification without a need for introduction of one or more breaks into any genetic material being modified.