Modified mRNA for Cytoplasmic Protein Production
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Solution Overview
Problem
Current methodologies for protein expression face challenges such as DNA integration into host cell genomic DNA, immunogenicity of mRNA, and inefficiencies in protein production, including low translation rates and immune response activation.
Innovation Solution
Development of modified mRNA (mmRNA) molecules with structural and chemical features that optimize formulation and delivery, enhance expression rates, and minimize immune response, such as pseudouridine and 5-methyl-cytosine modifications, to improve protein production and localization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If DNA is introduced into host cells for protein expression, then protein production is achieved, but DNA may integrate into host cell genomic DNA causing alterations and damage
Solution Approach 1:
The patent extracts the harmful integration step by using mRNA directly instead of DNA. The mRNA is introduced into the cytoplasm where it can be translated into protein without ever entering the nucleus or integrating into genomic DNA, thus eliminating the harmful effect while maintaining protein production capability
Solution Approach 2:
The patent uses mRNA as an intermediary molecule between the genetic information and protein production. Instead of using DNA that could integrate into the genome, mRNA serves as a temporary mediator that carries the coding information to ribosomes for translation, then degrades naturally without permanent genomic alteration
2Productivity
If mRNA is used for protein expression, then protein production is achieved, but mRNA is immunogenic and activates immune response
Solution Approach 1:
The patent applies parameter changes by chemically modifying the mRNA structure. Specific nucleosides in the mRNA are replaced with modified versions (such as pseudouridine, 5-methylcytidine) that change the molecular properties of the mRNA, reducing its recognition by immune sensors while maintaining its ability to be translated into protein
Solution Approach 2:
The patent creates a composite mRNA structure by combining modified nucleosides with standard nucleotides. This composite approach allows the mRNA to retain its functional properties for protein synthesis while incorporating modified components that reduce immunogenicity and increase stability
3Productivity
If standard mRNA is used for protein expression, then protein production occurs, but translation rates are low and protein concentrations are insufficient
Solution Approach 1:
The patent optimizes translation rates by changing parameters of the mRNA structure, including modifying the 5' cap structure, optimizing the 5' and 3' untranslated regions, and adjusting codon usage. These parameter changes enhance ribosome binding efficiency and translation initiation, leading to higher protein production rates and concentrations
4Productivity
If mRNA is introduced into cells, then protein expression is achieved, but mRNA degradation occurs rapidly reducing expression duration
Solution Approach 1:
The patent changes the chemical parameters of mRNA by incorporating modified nucleosides that are resistant to nucleases. These modifications alter the molecular structure of the mRNA backbone and bases, making the molecule more stable and resistant to degradation by cellular enzymes, thereby extending the duration of protein expression
Solution Approach 2:
The patent transforms the disposable nature of mRNA from a limitation to an advantage by using modified mRNA that degrades in a controlled manner. The modified mRNA maintains high expression rates during its lifetime and then degrades cleanly without persistent genomic integration, providing temporary but intense protein production suitable for therapeutic applications
Data Source
AI summary
The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.