Novel 5′-UTR Element Enhances mRNA Translation Efficiency
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Solution Overview
Problem
Current nucleic acid drugs face challenges in drug delivery and druggability, with mRNA vaccines and therapies requiring improved 5′-untranslated region (UTR) elements for enhanced protein expression and stability.
Innovation Solution
A novel artificial 5′-untranslated region element is developed by operably linking a transcription-enhancing sequence, a Kozac sequence, and a full-length or fragment sequence of a naturally highly expressed gene's 5′-UTR, optimized to exclude TOP motifs and uORFs, and incorporating a non-structural sequence that prevents secondary structure formation, enhancing mRNA translation efficiency and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If natural 5′-UTR sequences are used, then mRNA stability is improved, but translation efficiency is limited
Solution Approach 1:
The patent creates a composite 5′-UTR element that combines multiple functional sequences: a transcription-enhancing sequence (AATAA), a Kozac sequence (GCCACC), and an optimized fragment from a naturally highly expressed gene. This composite structure integrates the stability-providing characteristics of natural sequences with translation-enhancing elements, achieving both mRNA stability and high translation efficiency that neither component could provide alone.
2Stability of the object's composition
If full-length natural 5′-UTR sequences are used, then mRNA stability is improved, but expression complexity increases
Solution Approach 1:
The patent extracts and utilizes only the essential functional fragment of the natural 5′-UTR sequence that provides stability, while deliberately excluding non-essential elements such as TOP motifs and uORFs that would add complexity without providing proportional benefit. This extracted fragment approach maintains mRNA stability while simplifying the overall sequence design.
Solution Approach 2:
The 5′-UTR is segmented into distinct functional modules: a transcription-enhancing AATAA sequence, a Kozac GCCACC sequence for ribosome binding, and an optimized fragment from the natural gene. This segmentation allows each element to perform its specific function independently, reducing overall complexity while maintaining stability and translation efficiency.
3Reliability
If TOP motifs and uORFs are included, then natural sequence fidelity is maintained, but translation efficiency decreases
Solution Approach 1:
The patent identifies that TOP motifs and uORFs, while part of natural sequences, actually reduce translation efficiency. By deliberately excluding these elements, the invention converts a potential harm (reduced translation) into a benefit (enhanced expression efficiency), while maintaining reliability through the use of transcription-enhancing AATAA sequences and proper Kozac sequences that ensure faithful translation initiation.
Data Source
AI summary
A 5′-untranslated region element includes the full-length sequence of the 5′-untranslated region of a naturally highly expressed gene; or a fragment sequence of the 5′-untranslated region of a naturally highly expressed gene, wherein the fragment sequence of the 5′-untranslated region does not comprise a TOP motif site (e.g., CTTTT) or a uORF sequence (AUG upstream of an open reading frame); or a fragment sequence of the 5′-untranslated region of a naturally highly expressed gene, a non-structural sequence, a transcription-enhancing sequence, and a Kozac sequence, wherein the fragment sequence of the 5′-untranslated region lacks a TOP motif site or a uORF sequence. The 5′-untranslated region element of the present application makes nucleic acid molecules having extremely high protein expression performance and expression efficiency. The element is usable for constructing artificial nucleic acid molecules with high expression performance and has excellent application prospects in the development of nucleic acid therapeutic drugs.