5′/3′ UTR Sequence Design for Stable mRNA Expression
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Solution Overview
Problem
Existing mRNA molecules are unstable and prone to degradation, limiting their effectiveness in gene therapy and other applications due to insufficient stability and protein expression levels.
Innovation Solution
A specific combination of 5′ UTR and 3′ UTR sequences, selected from predefined sequences (SEQ ID NOs: 1-36), enhances mRNA stability and translation efficiency, thereby increasing protein expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If conventional mRNA molecules are used, then the basic translation function is achieved, but the mRNA is unstable and easy to be degraded by RNAase
Solution Approach 1:
The patent segments the mRNA molecule into functional regions (5' UTR, CDS, 3' UTR) and optimizes each segment independently. The 5' UTR is designed to protect against 5' exonucleases while the 3' UTR provides stability against 3' exonucleases and enhances translation. This segmentation allows targeted improvement of stability without compromising translation function.
Solution Approach 2:
The patent changes sequence parameters of the UTR regions, specifically selecting 5' UTR sequences with particular GC content and secondary structure properties, and 3' UTR sequences with specific length and composition. These parameter changes optimize the mRNA for both stability and translation efficiency, resolving the contradiction between stability and functional reliability.
2Stability of the object's composition
If mRNA stability is improved through UTR sequences, then degradation resistance increases, but the protein expression level may be limited without optimized translation elements
Solution Approach 1:
The patent merges the stabilizing function of the 3' UTR with the translation-enhancing function of the 5' UTR into a single mRNA molecule. The 5' UTR contains elements that promote ribosome binding and translation initiation, while the 3' UTR provides stability. This merging allows simultaneous achievement of both stability and high protein expression levels.
Solution Approach 2:
The patent applies local quality optimization by giving different functional characteristics to different regions: the 5' UTR is optimized for translation initiation with specific secondary structures and GC content, while the 3' UTR is optimized for stability with specific sequence composition and length. This local optimization resolves the contradiction between stability and productivity.
3Productivity
If 5' UTR and 3' UTR sequences are selected to increase translation efficiency, then protein expression increases, but the complexity of sequence selection and optimization increases
Solution Approach 1:
The patent identifies universal 5' UTR and 3' UTR sequences that can enhance translation efficiency across multiple genes and cell types. The 5' UTR sequences (such as those from human growth hormone or interferon alpha) and 3' UTR sequences (such as those from beta-globin or albumin) have been shown to work universally, reducing the need for gene-specific optimization and simplifying the selection process.
Solution Approach 2:
The patent uses copied and validated UTR sequences from well-characterized genes with known high expression patterns. By copying proven functional elements from genes like human growth hormone, interferon alpha, beta-globin, and albumin, the patent avoids de novo optimization complexity while achieving high protein expression levels across different applications.
Data Source
AI summary
The present disclosure relates to a UTR molecule and use thereof. Specifically, the present disclosure relates to an mRNA molecule comprising 5′ UTR and 3′ UTR. The UTR molecule can significantly increase mRNA expression level.