CYBA UTRs Enhance mRNA Translation Efficiency
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Solution Overview
Problem
Current mRNA therapeutics face challenges with strong immunogenicity and limited stability, particularly in terms of translation efficiency, which affects dosing and bioavailability of encoded proteins.
Innovation Solution
Incorporating specific untranslated regions (UTRs) derived from the human cytochrome b-245 alpha polypeptide (CYBA) gene, such as the 5' and 3' UTRs, into mRNA molecules to enhance translation efficiency by flanking the coding sequence, thereby improving protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If conventional mRNA is used for therapeutics, then the mRNA can be directly translated into protein in the cytoplasm without nuclear transport, but the mRNA exhibits strong immunogenicity and limited stability
Solution Approach 1:
The patent modifies the mRNA structure by replacing specific nucleotides (uridine with 5-methyluridine, cytidine with 5-methylcytidine) and incorporating modified nucleosides throughout the mRNA sequence. These parameter changes in molecular composition reduce immunogenicity and enhance stability while preserving the direct cytoplasmic translation capability
Solution Approach 2:
The patent creates a composite mRNA structure combining modified nucleosides (5-methyluridine, 5-methylcytidine) with standard nucleotides in specific ratios. This composite approach integrates the immunomodulatory properties of modified nucleosides with the translational functionality of conventional mRNA, achieving both reduced immunogenicity and improved stability
2Reliability
If mRNA stability is increased through chemical modifications, then immunogenicity is reduced, but translation efficiency may be affected
Solution Approach 1:
The patent applies different modification strategies to different regions of the mRNA molecule. The 5' UTR contains specific sequences (including Kozak consensus sequences) optimized for translation initiation, while the coding region incorporates modified nucleosides at controlled frequencies (20-80% replacement). The 3' UTR includes stability-enhancing elements like poly-A tails and stem-loop structures. This spatial differentiation of modification density and type preserves translation efficiency while achieving enhanced stability
Solution Approach 2:
The patent employs partial replacement of nucleotides with modified versions rather than complete replacement. Specifically, 20-80% of uridine is replaced with 5-methyluridine and 20-80% of cytidine is replaced with 5-methylcytidine, with optimal performance observed at intermediate levels. This partial action approach maintains sufficient recognition by cellular translation machinery while achieving the desired stability and reduced immunogenicity
3Productivity
If translation efficiency is increased to improve protein expression, then dosing intervals can be reduced, but mRNA stability may be compromised
Solution Approach 1:
The patent merges multiple functional elements into a single integrated mRNA design: 5' cap structure for translation initiation, optimized 5' UTR with Kozak sequences for efficient ribosome binding, coding region with modified nucleosides for stability, and 3' UTR with poly-A tail and stem-loop structures for enhanced half-life. This combination achieves both high translation efficiency and extended duration of action simultaneously
Data Source
AI summary
Described is an RNA molecule comprising (a) a coding region coding for a polypeptide; and (b) upstream of said coding region one or more UTR(s) comprising the sequence as shown in SEQ ID NO:1 or a sequence which shows 1 to 4 substitutions in comparison to SEQ ID NO:1 and which results in an RNA molecule having the same or a higher translation efficiency as an RNA molecule comprising an UTR comprising SEQ ID NO:1; and/or (c) downstream of said coding region one or more UTR(s) comprising the sequence as shown in SEQ ID NO:2 or a sequence which shows 1 to 7 substitutions in comparison to SEQ ID NO:2 and which results in an RNA molecule having the same or a higher translation efficiency as an RNA molecule comprising an UTR comprising SEQ ID NO:2; wherein said polypeptide encoded by said coding region is not a cytochrome b-245 alpha polypeptide (CYBA). Moreover, described is a nucleic acid molecule encoding the RNA molecule according to the present invention. Further, described is a vector comprising the nucleic acid molecule according to the present invention and to a host cell comprising the vector according to the present invention. Further, described is a pharmaceutical composition comprising the RNA molecule according to the present invention and optionally a pharmaceutically acceptable carrier. Moreover, described is a kit comprising the RNA molecule according to the present invention. Finally, described is the use of one or more UTR(s) as defined in (b) and/or one or more UTR(s) as defined in (c) for increasing the efficiency of translating a coding region of an RNA molecule into a polypeptide or a protein encoded by said coding region.