Structured Poly(A) Tail mRNA for Stability and Protein Expression
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Solution Overview
Problem
Current mRNA-based therapeutic preparations face challenges in maintaining stability and achieving high protein expression levels due to rapid degradation by ribonucleases, with limited optimization of the poly(A) tail structure.
Innovation Solution
A nucleic acid construct is developed with a poly(A) tail forming secondary or tertiary structures such as stem-loop, bulged stem, or pseudoknot structures using complementary binding sequences to enhance intracellular stability and protein expression, without interfering with the untranslated region (UTR) or coding sequence (CDS).
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If a simple poly(A) tail is used, then the mRNA structure is simple and easy to manufacture, but the mRNA is rapidly degraded by ribonucleases and has low stability
Solution Approach 1:
The patent changes the structural parameters of the poly(A) tail by introducing secondary structures (stem-loop, bulged stem, pseudoknot) formed through complementary binding sequences. This transforms the simple single-stranded poly(A) tail into complex structured configurations that resist ribonuclease degradation while maintaining manufacturability through in vitro transcription
Solution Approach 2:
The patent creates a composite structure by combining the poly(A) tail with complementary binding sequences that form secondary structures. This composite configuration integrates both the protective function of poly(A) tails and the stability-enhancing properties of structured RNA elements, achieving high stability without compromising ease of manufacture
2Reliability
If the poly(A) tail is optimized for stability, then mRNA stability improves, but protein expression levels remain limited
Solution Approach 1:
The patent applies local quality by creating specific secondary structures at particular regions within the poly(A) tail. Different structural configurations (stem-loop, bulged stem, pseudoknot) are positioned strategically to provide localized stability enhancement while maintaining overall translational efficiency. The complementary binding sequences are designed to form structures that protect against degradation without interfering with ribosome binding and translation
Solution Approach 2:
The patent introduces dynamic characteristics to the poly(A) tail structure through complementary binding sequences that can form reversible secondary structures. These dynamic structures allow the mRNA to maintain flexibility in translation while providing stability protection, enabling both high stability and high protein expression levels simultaneously
3Reliability
If complementary binding sequences are inserted into the poly(A) tail, then secondary structures form to enhance stability, but the structure becomes more complex
Solution Approach 1:
The patent segments the poly(A) tail into distinct functional regions by inserting complementary binding sequences at specific positions. This segmentation creates modular secondary structures (stem-loop, bulged stem, pseudoknot) that can be independently optimized while maintaining overall structural coherence. The segmented approach allows systematic design of stability-enhancing structures without overwhelming complexity
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 construct significantly enhances mRNA stability and protein expression levels by forming stable secondary or tertiary structures, ensuring consistent high efficiency across various mRNA forms.
Implementation Method 1
a poly(A) tail having a secondary or tertiary structure
Data Source
Figure 1A~1B
Figure 2
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AI summary
The present invention relates to a novel nucleic acid construct having improved stability and protein expression level and uses thereof and, more specifically, to a nucleic acid construct comprising: a coding region encoding a polypeptide or protein; and a poly(A) tail having a secondary or tertiary structure, and a pharmaceutical composition for vaccine or gene therapy, comprising the nucleic acid construct. A nucleic acid construct platform according to the present invention exhibits more improved stability and protein expression rate in cells without interfering with CDS and UTR sequences and the like, and thus can be generally and effectively used in gene therapy, vaccine fields, and the like.