Artificial Nucleic Acid Molecules With TOP-Gene UTRs for Stable Expression
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Solution Overview
Problem
Current gene therapy and genetic vaccination methods using nucleic acid molecules face challenges such as genomic integration risks, limited expression levels, and instability of RNA, particularly due to degradation by RNases, which affect the efficacy of protein production and immune response induction.
Innovation Solution
Development of artificial nucleic acid molecules, including a 5′ UTR element derived from the 5′ UTR of a TOP gene, optionally with a histone stem-loop and a 3′ UTR element, to enhance stability and translational efficiency, utilizing a G/C-rich sequence and a poly(A) tail for improved mRNA expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional nucleic acid molecules are used for gene therapy and genetic vaccination, then the methods can be applied to treat diseases and induce immune responses, but the RNA is unstable and degraded by RNases, limiting efficacy
Solution Approach 1:
The patent modifies the nucleic acid molecule by changing sequence parameters - specifically incorporating a 5' UTR element derived from TOP genes and optimizing the coding sequence to increase GC content. These parameter changes enhance mRNA stability and resistance to RNase degradation, directly addressing the RNA instability problem while maintaining therapeutic function.
Solution Approach 2:
The invention creates a composite nucleic acid structure combining multiple functional elements: the 5' UTR element from TOP genes, the optimized coding sequence with enhanced GC content, and the 3' UTR element. This composite structure synergistically improves mRNA stability and translational efficiency while resisting degradation.
2Productivity
If nucleic acid molecules are used to express proteins, then protein production can be achieved, but the expression levels are limited
Solution Approach 1:
The patent optimizes translation parameters by increasing the GC content of the coding sequence and incorporating specific 5' UTR elements from TOP genes known to enhance translational efficiency. These parameter modifications directly increase protein production levels while maintaining mRNA stability.
Solution Approach 2:
The invention uses the 5' UTR element from TOP genes as a template or model to design the 5' UTR region of the therapeutic mRNA. This copying of a proven functional element enhances translational efficiency and protein expression levels in the therapeutic context.
3Duration of action of stationary object
If genomic integration is used to ensure long-term expression, then protein production is sustained, but the risk of harmful genomic integration occurs
Solution Approach 1:
The patent extracts and utilizes only the necessary functional elements (5' UTR from TOP genes, optimized coding sequence, 3' UTR) without requiring integration into the host genome. This extraction approach enables long-term expression through mRNA stability and translational enhancement while avoiding the harmful effects of genomic integration.
Solution Approach 2:
The invention employs a non-integrating, transient mRNA approach that achieves sustained expression through molecular optimization rather than permanent genomic insertion. This disposable mRNA strategy avoids the risks of genomic integration while maintaining adequate expression duration for therapeutic effect.
Data Source
AI summary
The invention relates to an artificial nucleic acid molecule comprising at least one 5′ UTR element which is derived from a TOP gene, at least one open reading frame, and preferably at least one histone stem-loop. Optionally the artificial nucleic acid molecule may further comprise, e.g. a poly(A)sequence, a poyladenylation signal, and/or a 3′ UTR. The invention further relates to the use of such an artificial nucleic acid molecule in gene therapy and/or genetic vaccination.