β-Catenin 3′UTR Sequence Optimization for Stable Protein Secretion
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Solution Overview
Problem
Existing technologies fail to effectively enhance the translation efficiency and promote the movement of target proteins into the cytoplasm or cellular membrane for stable secretion, particularly for β-catenin mRNA, lacking studies on the impact of 3′UTR isoforms on protein expression and localization.
Innovation Solution
An mRNA molecule comprising a coding region with a 5′UTR and a specific 3′UTR fragment of β-catenin mRNA, optimized by deletions of intron 15 and exon 16A, enhances translation efficiency and promotes protein movement into the cytoplasm or cellular membrane.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional mRNA sequences are used, then basic protein expression occurs, but translation efficiency and protein localization to cytoplasm/cell membrane are insufficient
Solution Approach 1:
The patent changes the nucleotide sequence parameters of the 3'UTR region by deleting specific sequences (intron 15 and exon 16A) to optimize translation efficiency and protein localization. This sequence optimization directly addresses the contradiction by modifying molecular parameters to achieve both high productivity and reliable secretion.
Solution Approach 2:
The patent segments the 3'UTR region into specific functional elements, identifying and removing inhibitory segments (intron 15 and exon 16A) while retaining beneficial segments. This segmentation allows precise control over translation efficiency and protein localization, resolving the contradiction between expression level and secretion stability.
2Productivity
If full-length 3'UTR isoforms are used, then mRNA stability is maintained, but protein expression efficiency and localization control are limited
Solution Approach 1:
The patent extracts and removes specific inhibitory elements (intron 15 and exon 16A sequences) from the full-length 3'UTR, creating a simplified yet highly efficient mRNA structure. This extraction resolves the contradiction by eliminating complexity-reducing elements while enhancing protein expression efficiency.
Solution Approach 2:
The patent applies local quality modification by specifically optimizing the 3'UTR region while maintaining other mRNA components. The localized sequence changes in the 3'UTR create specific functional properties that enhance translation and localization without requiring changes throughout the entire mRNA molecule, thus managing structural complexity efficiently.
3Ease of operation
If standard UTR sequences are used, then general protein production occurs, but targeted protein movement to cytoplasm and cell membrane is insufficient
Solution Approach 1:
The patent changes the sequence parameters of the 3'UTR to create specific localization signals that direct protein movement to the cytoplasm and cell membrane. These parameter changes in nucleotide sequence directly improve both localization efficiency and the quantity of secreted protein, resolving the contradiction between ease of operation and substance quantity.
Data Source
AI summary
The present invention relates to an mRNA including the UTR of polymorphic β-catenin. When used, the mRNA molecule, nucleic acid molecule, expression construct, and/or expression vector of the present invention can effectively enhance the expression efficiency of a target protein and extend the expression location of the target protein to the cytoplasm, thus allowing for stable extracellular secretion. In addition, the mRNA functions to regulate an expression level of exogenously introduced mRNA and thus can be utilized as an mRNA vaccine in the future.