Optimized mRNA for Genome Editing Reducing Immunogenicity
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Solution Overview
Problem
Existing genome editing approaches using RNA-guided DNA binding agents like CRISPR-Cas systems face inefficiencies and immunogenicity issues, leading to suboptimal cytokine responses upon administration.
Innovation Solution
Development of mRNA encoding RNA-guided DNA-binding agents with optimized codon usage, uridine content, and nucleotide composition to enhance editing efficiency and reduce immunogenicity, specifically targeting liver cells with improved translation efficiency and reduced cytokine elevation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing mRNA encoding RNA-guided DNA binding agents is used for genome editing, then genome editing function is achieved, but editing efficiency is insufficient and immunogenicity increases
Solution Approach 1:
The patent optimizes multiple parameters of the mRNA molecule including codon usage (using codons preferred in human cells), uridine content (reducing to minimum necessary levels), nucleotide composition ratios, and structural features to simultaneously improve translation efficiency and reduce immunogenicity. This involves changing the chemical and sequence parameters of the mRNA to achieve better performance
Solution Approach 2:
The patent applies different optimization strategies to different regions of the mRNA molecule, such as optimizing the 5' UTR and 3' UTR regions separately from the coding sequence, and using different codon optimization approaches for different segments of the open reading frame to maximize translation efficiency while minimizing immune recognition
2Productivity
If codon usage and nucleotide composition are optimized to improve translation efficiency, then editing efficiency increases, but complexity of mRNA design increases
Solution Approach 1:
The patent systematically optimizes multiple parameters including codon usage frequency, uridine content, adenine content, GC content, and nucleotide composition ratios. By establishing specific parameter ranges and optimization criteria, the complex design process is structured and manageable
Solution Approach 2:
The patent performs preliminary optimization of the mRNA sequence before synthesis, including codon optimization based on human cell preferences, adjustment of nucleotide composition, and design of optimized 5' and 3' UTR regions. This preliminary design work simplifies the overall development process by establishing optimized sequences before experimental validation
Data Source
AI summary
Compositions and methods for gene editing. In some embodiments, a polynucleotide encoding Cas9 is provided that can provide one or more of improved editing efficiency, reduced immunogenicity, or other benefits.