mRNA UTR Complementarity Layout for Higher Protein Expression
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Solution Overview
Problem
Existing mRNA constructs for protein expression in vaccines and therapeutics face challenges in optimizing translation efficiency and safety, particularly in regulating protein expression levels through the degree of complementarity between untranslated regions.
Innovation Solution
A nucleic acid construct is designed with partially complementary 5' and 3' untranslated regions (UTRs) that include non-complementary regions, with specific patterns and lengths to enhance protein expression levels, utilizing a method that calculates free energy changes and measures expression levels in cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the degree of complementarity between 5' UTR and 3' UTR is increased to enhance protein expression, then translation efficiency is improved, but the risk of forming secondary structures that may interfere with translation initiation increases
Solution Approach 1:
The patent applies local quality by creating specific non-complementary regions at critical positions within the UTR sequences. These localized non-complementary segments prevent secondary structure formation at the 5' end while maintaining complementarity in other regions to promote circularization and overall translation efficiency. This selective modification of specific regions rather than uniform complementarity resolves the contradiction between enhancing expression and maintaining reliable translation initiation.
2Productivity
If completely complementary 5' UTR and 3' UTR sequences are used to maximize circularization, then mRNA stability and translation efficiency are improved, but the complexity of sequence design and optimization increases
Solution Approach 1:
The patent segments the UTR sequences into distinct complementary regions and non-complementary regions with specific functional roles. This segmentation simplifies the design process by providing a modular framework where complementary segments promote circularization while non-complementary segments prevent unwanted secondary structures. The segmented approach reduces design complexity compared to attempting complete complementarity while maintaining high translation efficiency.
3Reliability
If non-complementary regions are introduced to prevent secondary structures, then translation initiation is improved, but the degree of complementarity and circularization efficiency decreases
Solution Approach 1:
The patent applies partial action by introducing non-complementary regions only at specific critical positions rather than throughout the entire UTR sequence. This partial modification is sufficient to prevent secondary structure formation and ensure translation initiation efficiency while maintaining adequate complementarity in other regions to promote circularization and high protein expression levels. The selective rather than complete approach resolves the contradiction between reliable initiation and high productivity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The nucleic acid construct achieves higher protein expression levels compared to conventional constructs, ensuring safety and efficiency in mRNA-based therapies.
Implementation Method 1
a 5' untranslated region (UTR) and a 3' UTR that are at least partially complementary to each other
Implementation Method 2
a degree of complementarity of the 3' UTR to the 5' UTR is greater than 75%
Data Source
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AI summary
The present disclosure provides a nucleic acid construct capable of regulating protein expression. Specifically, the present disclosure provides a nucleic acid construct including a 5' untranslated region (UTR) and a 3' UTR that are at least partially complementary to each other, wherein at least one of UTRs includes a non-complementary region and a complementary region with respect to the other UTR, and when all of the respective lengths of the non-complementary regions are of 1 base, a degree of complementarity of the 3' UTR to the 5' UTR is greater than 75%.