Hybridized Guide Nucleic Acids for Template-Based Gene Editing
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Solution Overview
Problem
Current gene editor systems face challenges in efficiently assembling guide nucleic acids for gene editing, particularly in synthesizing and hybridizing multiple RNA molecules, which can lead to complexity and reduced manufacturing quality.
Innovation Solution
The development of hybridized guide nucleic acids, comprising multiple polynucleotides that hybridize to form a functional guide nucleic acid, allowing for modular assembly and improved manufacturing quality, and enabling the use with template-based gene editors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple RNA molecules are synthesized and hybridized to form guide nucleic acids, then the functionality of gene editing is achieved, but the manufacturing complexity and error rates increase
Solution Approach 1:
The guide nucleic acid is divided into multiple separate polynucleotide components that can be synthesized independently and then hybridized together. This segmentation allows each component to be manufactured with higher quality control while maintaining the overall functionality of the complete guide nucleic acid structure.
Solution Approach 2:
Multiple separately synthesized polynucleotide components are combined through hybridization to form the complete functional guide nucleic acid. This merging approach enables modular assembly that reduces manufacturing complexity while preserving the reliability benefits of individual component synthesis.
2Ease of manufacture
If single guide RNA is used, then synthesis is simpler, but modifications cannot be made without affecting hybridization
Solution Approach 1:
The guide RNA is segmented into multiple polynucleotide components, allowing modifications to be introduced in one or more segments without affecting the hybridization interfaces. This enables versatile modifications while maintaining synthesis simplicity through modular construction.
Solution Approach 2:
The modular polynucleotide components can serve multiple functions - some segments provide hybridization capability while others accommodate modifications. This multi-functionality allows the system to maintain ease of manufacture while gaining adaptation flexibility for various editing applications.
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
This approach facilitates the assembly of guide nucleic acid libraries and improves manufacturing quality by simplifying the synthesis process, enhancing the efficiency of gene editing and reducing errors.
Implementation Method 1
The first and second hybridization sequences are complementary. The first and second hybridization sequences hybridizes to one another to form the hybridized guide nucleic acid.
Data Source
AI summary
Polynucleotides are engineered to hybridize to one another to form a functional guide nucleic acid having an editing template. The hybridized guide nucleic acids may be used with template-based gene editors to write an edit into a target genomic location using the editing template as a template for the edit.