IRES-Optimized Plasmid Vectors for Stable Circular mRNA Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing circular RNA technologies face challenges with size and limited internal ribosome entry site (IRES) activity, leading to inefficient protein expression and stability, particularly when applied in vivo.
Innovation Solution
Development of a plasmid vector for creating open reading frame-coding circular mRNA with optimized homology arms and IRES, resulting in a small backbone design that enhances translation efficiency and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If circular RNA is used to extend mRNA activity and stability, then protein expression duration is improved, but translation efficiency deteriorates due to size and limited IRES activity
Solution Approach 1:
The circular RNA molecule is segmented into functional modules including IRES elements, 5' and 3' UTRs, and coding regions. This modular segmentation allows optimization of each component's function while maintaining overall circular structure stability, thereby improving both translation efficiency and duration of action
Solution Approach 2:
The patent optimizes parameters such as IRES sequence selection, UTR length, and circularization efficiency to enhance both translation initiation and mRNA stability. By adjusting these parameters, the circular RNA achieves improved protein expression duration without sacrificing translation efficiency
2Reliability
If RNA circularization is performed to reduce exonuclease excretion and increase half-life, then stability is improved, but protein expression level deteriorates over time
Solution Approach 1:
IRES elements serve as intermediaries that facilitate ribosome binding and translation initiation on circular RNA. The patent incorporates optimized IRES sequences that mediate efficient translation while the circular structure provides stability, resolving the contradiction between long half-life and sustained protein expression levels
Solution Approach 2:
The circular RNA construct combines multiple functional elements (IRES, UTRs, coding sequences) into a composite molecular structure. This composite design integrates the stability benefits of circularization with the translation efficiency of viral IRES elements, maintaining high protein expression levels over extended periods
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The optimized plasmid vector and circular mRNA significantly improve protein expression in vivo, reducing the frequency of administration and potential side effects, and increasing patient access to medicines.
Implementation Method 1
During protein synthesis, ribosomes bind to mRNA to read the nucleotide sequences, which are then translated into proteins
Implementation Method 2
a plasmid vector for making an open reading frame-coding circular mRNA (ORF-coding circular mRNA), said vector comprising elements that are connected to each other and arranged in a following sequence including a 5′ homology arm connected to a 3′ group I intron fragment
Data Source
AI summary
The present invention relates to a plasmid vector for making an open reading frame-coding circular mRNA (ORF-coding circular mRNA) having a small backbone, optimized homology arms and internal ribosome entry site (IRES) that encodes an efficient open reading frame (ORF), resulting in highly efficient protein expression. The IRES has also been developed to promote the translation of proteins of interest, thus improving protein expression. The plasmid vector and ORF-coding circular mRNA according to this invention can overcome previous technical challenges due to its optimized constructs and small size, resulting in ease of delivery, stability in the cell, and significantly higher translation efficiency and protein expression in vivo. When applied in medicine or pharmaceutical preparations, the circular mRNA described in the present invention can potentially reduce the frequency of administration and/or doses required, resulting in fewer unwanted side effects and improved patient access to medicines.