In-Vitro Transcript mRNA Composition for Stable Vaccine Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing genetic therapies using DNA and RNA for gene expression face challenges such as genetic damage, limited protein expression, and inconsistent immune responses due to species-specific expression and antigen recognition, necessitating improved mRNA stability and translation activity.
Innovation Solution
Development of in-vitro transcript mRNA with 5′-UTR and 3′-UTR, a 5′ cap, and a poly(A) tail containing 20 to 400 adenines for stable protein expression in animal cells, using a template DNA for production and a pharmaceutical composition for vaccines.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If DNA is used for gene administration, then stability and ease of handling are improved, but genetic damage risk and limited protein expression occur
Solution Approach 1:
The patent extracts the gene of interest from DNA and uses only the RNA transcript for gene administration. By separating the transcription step (done in vitro) from the translation step (done in vivo), the patent eliminates the risk of DNA insertion into the genome while maintaining gene expression capability. The RNA is produced in vitro and directly delivered to cells, avoiding the need for DNA to integrate into cellular chromosomes.
Solution Approach 2:
The patent uses an in vitro transcribed RNA as an intermediary between the gene template and the cellular expression machinery. The RNA serves as a temporary messenger that carries genetic information from the in vitro synthesized template to the cellular translation apparatus, enabling protein expression without requiring long-term genomic integration.
2Ease of operation
If DNA is used for gene administration, then gene delivery is achieved, but protein expression level is limited due to transcription factor dependency
Solution Approach 1:
The patent performs transcription in advance (in vitro) to produce the RNA transcript before delivery to cells. By completing the transcription step externally using purified RNA polymerase and nucleotide substrates, the patent eliminates the need for cellular transcription factors and chromatin accessibility, thereby maximizing translation efficiency and protein expression levels upon cellular delivery.
3Object-generated harmful factors
If RNA is used for gene administration, then direct cytoplasmic expression and no genetic damage occur, but rapid enzymatic degradation limits duration of action
Solution Approach 1:
The patent optimizes RNA parameters including adding a 5' cap structure and poly(A) tail to enhance stability and translation efficiency. These structural modifications protect the RNA from enzymatic degradation while maintaining its ability to be translated into protein, thereby extending the functional duration of the RNA in cellular environments.
4Quantity of substance
If large amount of RNA is produced in vitro, then sufficient gene expression is achieved, but production cost and complexity increase
Solution Approach 1:
The patent uses a DNA template to in vitro transcribe multiple copies of the RNA transcript. A single DNA template can produce numerous RNA molecules through in vitro transcription, enabling large-scale RNA production without requiring complex biological systems. This copying approach allows RNA mass production in simplified facilities compared to traditional biological reactor methods.
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 in-vitro transcript mRNA platform enables high and stable protein expression, inducing effective immune responses and facilitating rapid vaccine production without the need for biological reactors, suitable for genetic vaccines against autoimmune, infectious, and cancer-related diseases.
Implementation Method 1
the in-vitro transcript mRNA for intracellular expression of a gene of interest
Implementation Method 2
5' cap linked to 5'-UTR, and a poly(A) tail containing 20 to 400 adenines linked to 3'-UTR
Data Source
AI summary
The present invention relates to an RNA in-vitro transcript mRNA for intracellular expression of a gene of interest and a pharmaceutical composition comprising same for vaccines. When injected into animal cells, the in-vitro transcript mRNA including the gene of interest according to the present invention allows the protein of interest to be expressed in the animal cell in large quantities, and, as such, can be used as a gene vaccine against autoimmune diseases, infectious diseases, cancer- or tumor-related diseases, inflammatory diseases, and so on.