Structure-engineered guide RNA for CRISPR delivery
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current genome editing methods using CRISPR/Cas technology face challenges in safely and efficiently delivering components like guide RNA and Cas9 into cells due to toxicity and immunogenicity issues with viral methods and inefficiencies with non-viral approaches.
Innovation Solution
Structure-engineered guide RNA molecules with extension sequences, including crRNA and tracrRNA, are developed to enhance delivery and functionality, comprising self-hybridizing sequences that form structured RNA extensions, which can be used alone or with a vehicle to facilitate gene editing by contacting cells with these modified RNAs and an RNA-guided endonuclease.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral methods are used to deliver CRISPR/Cas components into cells, then delivery efficiency is improved, but toxicity and immunogenicity increase
Solution Approach 1:
The patent uses non-viral delivery vehicles (such as lipid nanoparticles, polymers, or other non-viral carriers) as intermediaries to transport CRISPR/Cas components into cells. These non-viral vectors serve as mediators that achieve efficient cellular delivery without the toxicity and immunogenicity associated with viral vectors, thus resolving the contradiction between delivery efficiency and safety
2Object-affected harmful factors
If non-viral methods are used to deliver CRISPR/Cas components into cells, then safety is improved, but delivery efficiency decreases
Solution Approach 1:
The patent modifies parameters of non-viral delivery systems, including optimizing the chemical composition, physical properties (such as particle size, surface charge), and structural characteristics of non-viral vectors. By adjusting these parameters, the delivery efficiency of non-viral methods is enhanced to match or approach viral-level efficiency while maintaining safety advantages
3Productivity
If guide RNA is extended with additional sequences, then cellular uptake and interaction with CRISPR/Cas system are improved, but molecule complexity increases
Solution Approach 1:
The patent divides the guide RNA molecule into distinct functional segments: the essential guide sequence for target recognition and binding, and additional extension sequences that enhance cellular uptake and stability. This segmentation allows the core functional element to remain simple while adding specialized modules that improve overall performance without compromising the fundamental guide RNA structure
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 modified guide RNA molecules improve the efficiency and safety of gene editing by enhancing cellular uptake and interaction with the CRISPR/Cas system, allowing precise targeting and editing of genes while minimizing toxicity and immunogenicity concerns.
Implementation Method 1
comprising self-hybridizing sequences that form structured RNA extensions
Data Source
AI summary
The invention provides a modified guide RNA comprising an extension sequence, as well as related compositions and methods.