Modified mRNA for Reduced Immunogenicity and Faster Protein Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methodologies for protein expression, such as DNA-based approaches, face challenges including integration into host genomic DNA, multiple processing steps, and difficulty in achieving efficient and sustained protein production, especially in primary cells or modified cell lines, leading to lag times and variable expression rates.
Innovation Solution
Development of modified messenger RNA (mRNA) molecules incorporating specific modified nucleosides, such as 5-methylcytosine, N6-methyladenosine, and 2′-O-methylated nucleosides, which are designed to enhance translation efficiency and reduce immunogenicity, allowing for direct protein production in mammalian cells with improved expression rates and reduced immune response.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If DNA-based approaches are used for protein expression, then genetic material can be inherited by daughter cells, but integration into host genomic DNA causes alterations and damage to host cell genomic DNA
Solution Approach 1:
The patent extracts the protein expression function from DNA-based approaches and implements it directly through mRNA molecules. By using mRNA instead of DNA, the invention eliminates the need for genomic integration while maintaining the ability to produce proteins, thus avoiding damage to host cell genomic DNA while still achieving sustained protein expression through repeated administrations
Solution Approach 2:
The patent introduces mRNA as an intermediary between genetic information and protein production. This intermediary approach allows protein expression without requiring DNA integration into the host genome, thereby avoiding the harmful effects of genomic DNA alteration while maintaining reliable protein production capability
2Reliability
If DNA-based approaches are used for protein expression, then genetic material can be introduced into cells, but multiple processing steps create lag times before protein generation
Solution Approach 1:
The patent extracts the essential protein expression function from the complex DNA-based multi-step process and implements it directly through mRNA. By bypassing nuclear transport, transcription, and other intermediate steps, the invention achieves direct translation of mRNA to protein, significantly reducing lag time while maintaining reliable protein production
Solution Approach 2:
Instead of following the natural central dogma flow (DNA → RNA → Protein), the patent inverts the approach by directly introducing mRNA (the intermediate product) into the cytoplasm for immediate translation. This inversion eliminates unnecessary processing steps and accelerates protein generation while preserving production capability
3Reliability
If DNA expression is introduced into cells, then genetic material can be delivered, but DNA enters cells but is not expressed or not expressed at reasonable rates or concentrations
Solution Approach 1:
The patent extracts the protein expression function from DNA and implements it directly through mRNA molecules. By using mRNA as the delivery vehicle, the invention ensures immediate availability for translation without relying on complex nuclear import and transcription machinery, thereby achieving both successful delivery and high expression rates
Solution Approach 2:
The patent changes the molecular form from DNA to mRNA, fundamentally altering the expression parameters. mRNA molecules are directly translatable in the cytoplasm, eliminating rate-limiting steps such as nuclear transport and transcription initiation, thereby achieving reasonable to high expression rates while maintaining delivery capability
4Productivity
If unmodified mRNA is used for protein production, then protein can be produced in mammalian cells, but immunogenicity limits sustained expression especially in primary cells or modified cell lines
Solution Approach 1:
The patent applies local quality modification by introducing specific modified nucleosides at particular positions within the mRNA sequence. These localized modifications (such as pseudouridine, 5-methylcytidine, or N6-methyladenosine) reduce immunogenicity at specific sites without compromising the overall protein production efficiency, enabling sustained expression in primary and modified cell lines
Solution Approach 2:
The patent creates composite mRNA molecules combining standard nucleosides with modified nucleosides in specific patterns. This composite structure reduces immunogenic recognition while maintaining translation efficiency, thereby achieving both high productivity and reduced immunogenicity simultaneously through the synergistic effect of different nucleoside components
Data Source
AI summary
This invention provides messenger RNA (mRNA) molecules comprising an open reading frame that encodes a protein of interest, wherein said modified RNA comprises a modified nucleoside selected from the group consisting of: (I), (II), and (III), gene therapy vectors comprising same, methods of synthesizing same, and methods for gene replacement, gene therapy, gene transcription silencing, and the delivery of therapeutic proteins to tissue in vivo, comprising the molecules. The present invention also provides methods of reducing the immunogenicity of mRNA molecules.