Artificial Nucleic Acid Molecules for Stable Protein Expression
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Solution Overview
Problem
Current gene therapy and genetic vaccination methods face challenges with the stability and expression efficiency of nucleic acid molecules, including DNA's risk of genomic integration and limited expression levels, and RNA's instability, which affects the efficacy of protein production for therapeutic and immunological applications.
Innovation Solution
The development of artificial nucleic acid molecules incorporating a 5'UTR element derived from a TOP gene, optionally a histone stem-loop, a 3'UTR element, a poly(A) sequence, and/or a polyadenylation signal, designed to enhance protein production and translational efficiency by stabilizing the mRNA and promoting efficient translation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If natural RNA molecules are used for gene therapy and genetic vaccination, then the risk of genomic integration and anti-DNA antibody generation is minimized, but the RNA molecules are rapidly degraded by RNases leading to poor stability and limited expression
Solution Approach 1:
The patent modifies the chemical structure of RNA molecules by incorporating modified nucleotides (e.g., pseudouridine, 5-methylcytidine, N1-methyladenosine) to change parameters of the RNA molecule. These modifications reduce recognition by RNases while maintaining translation capability, thereby improving stability without compromising safety
Solution Approach 2:
The invention creates composite nucleic acid molecules combining natural RNA with chemically modified nucleotides. This composite structure leverages the safety profile of natural RNA while adding the stability benefits of modified nucleotides, achieving both improved reliability and stability simultaneously
2Productivity
If conventional nucleic acid molecules are used, then the basic transcription and translation machinery is sufficient, but the translational efficiency is limited and protein production is insufficient for therapeutic effects
Solution Approach 1:
The patent divides the nucleic acid molecule into distinct functional segments: 5'UTR with specific sequences (e.g., CUG-5' terminal oligopyrimidine tract), coding region with optimized codons, and 3'UTR with stability elements. Each segment is engineered independently to contribute specific functions, improving overall protein production through coordinated action of optimized parts
Solution Approach 2:
The invention applies local optimizations at specific positions in the nucleic acid sequence. The 5'UTR contains specific sequences (CUG-5' terminal oligopyrimidine tract, UG-rich region) that locally enhance translation initiation, while the 3'UTR contains stability elements that locally protect against degradation. These localized improvements collectively boost protein production without requiring complete redesign of the entire molecule
Data Source
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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.