Modified RNA Nucleosides for Low-Immunogenic Protein Expression
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Solution Overview
Problem
Existing in vitro-synthesized RNA molecules, such as mRNA, oligoribonucleotides, and polyribonucleotides, exhibit high immunogenicity, leading to significant cytokine secretion when interacting with dendritic cells, which can trigger immune responses.
Innovation Solution
Incorporation of pseudouridine (Ψ) or 1-methylpseudouridine (m1Ψ) into the RNA molecules, along with a poly-A tail, m7GppG cap, and cap-independent translational enhancers, reduces immunogenicity by measuring decreased cytokine secretion in murine or human dendritic cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If in vitro-synthesized RNA molecules are used, then protein expression can be achieved, but immunogenicity increases leading to cytokine secretion
Solution Approach 1:
The patent applies parameter changes by incorporating modified nucleosides (pseudouridine Ψ, 1-methylpseudouridine m1Ψ, 5-methylcytidine m5C) into the RNA molecules. These chemical modifications change the nucleoside parameters of the RNA, reducing its immunogenicity while maintaining protein expression capability. The modified RNA molecules show decreased cytokine secretion in dendritic cells compared to unmodified RNA.
2Productivity
If unmodified in vitro-synthesized RNA is used, then translation can occur, but immune response is triggered
Solution Approach 1:
The patent modifies the RNA molecules by incorporating pseudouridine (Ψ), 1-methylpseudouridine (m1Ψ), and/or 5-methylcytidine (m5C) at specific positions within the RNA sequence. These parameter changes in the nucleoside structure reduce the RNA's ability to trigger cytokine secretion in dendritic cells while preserving its translation efficiency for protein expression.
Data Source
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AI summary
This invention provides RNA, oligoribonucleotide, and polyribonucleotide molecules comprising pseudouridine or a modified nucleoside, 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 RNA, oligoribonucleotide, and polyribonucleotide molecules.