mRNA Transcription Vector UTR Design for Stable Protein Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing mRNA vaccines face challenges in achieving high stability, translation efficiency, and immunogenicity due to innate immune responses and low expression of antigen proteins, limiting their effectiveness in therapeutic and vaccine applications.
Innovation Solution
An mRNA transcription vector is developed with a promoter region recognized by RNA polymerase and a gene construct comprising a 5'-untranslated region (5'-UTR), open reading frame (ORF), and 3'-untranslated region (3'-UTR) to enhance mRNA expression and immunogenicity, utilizing a specific nucleotide sequence with 90% or more identity to SEQ ID NO: 1, 2, and 3, respectively.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-generated harmful factors
If naked mRNA is used to induce immune response, then immunogenicity is enhanced, but transcription signaling is inhibited due to TLR agonist stimulation
Solution Approach 1:
The mRNA molecule is segmented into distinct functional regions: 5'-UTR (nucleotides 1-39 from SEQ ID NO: 1), ORF (nucleotides 40-381 encoding SARS-CoV-2 spike protein), and 3'-UTR (nucleotides 382-421 from SEQ ID NO: 2). This segmentation allows independent optimization of each region for translation efficiency while maintaining immunogenicity through specific sequence features.
Solution Approach 2:
The patent modifies specific nucleotide sequences and their lengths to optimize both translation and immunogenicity. The 5'-UTR uses a specific sequence from SEQ ID NO: 1 with defined length, the ORF encodes specific spike protein variants (including Omicron BA.1, BA.2, BA.3, and BA.4/5), and the 3'-UTR uses sequences from SEQ ID NO: 2 with controlled repetition patterns. These parameter changes enable high translation efficiency while preserving immune activation capabilities.
2Stability of the object's composition
If mRNA stability is increased for therapeutic application, then translation efficiency decreases due to innate immune responses
Solution Approach 1:
Different regions of the mRNA molecule are assigned different functional qualities: the 5'-UTR is optimized for ribosome binding and translation initiation with specific sequence from SEQ ID NO: 1, the ORF region is optimized for protein coding accuracy, and the 3'-UTR is optimized for stability with repetitive sequences from SEQ ID NO: 2. This local optimization allows each region to perform its specific function at maximum efficiency.
Solution Approach 2:
The mRNA construct combines multiple sequence elements into a composite structure: the 5'-UTR from SEQ ID NO: 1, the ORF encoding spike protein, and the 3'-UTR from SEQ ID NO: 2 with repetitive patterns. This composite design integrates stability-enhancing features with translation-optimizing features, creating an mRNA molecule that simultaneously achieves both properties.
Data Source
AI summary
The present application relates to mRNA for protein expression and a template therefor, and provides an mRNA transcription vector including a gene construct according to an embodiment, a method of producing an mRNA molecule including the processes of performing in vitro transcription using the mRNA transcription vector, an mRNA molecule prepared by the method, and an immunogenic composition including the mRNA molecule as an active ingredient.